產品資訊 > 人類 miRNA 晶片 Human miRNA OneArray® v7

 
    人類 miRNA 晶片 v7 延續 v6 的設計原理, 參考 Sanger miRBase 資料庫 release 21 的序列進行探針設計, 並以自有的高速佈放技術進行晶片生產。v7 版晶片於 201511月上市。
 
 
 
人類 miRNA 晶片 v7 探針內容
  學名 Homo sapiens
  常用中文名稱 人類
  探針代表號 hsa
  常用英文名稱 Human
  miRNA 探針數 2,548
  總所對應的成熟 miRNA 2,588
  miRNA 探針重複數 3
  控制探針數 124
  總探針數 7,768


  基 因 探 針

人類 miRNA 晶片 v7 是參考 Sanger miRBase 資料庫 release 21 的序列進行探針設計, 共有 2,548 個序列,可對應到 2,588 個成熟的 miRNA, 在探針設計的過程,依據各序列的特性,進行嚴格 Tm 值的把關及調整。 在每一晶片上,每一個 miRNA 序列具有 3 個重覆的探針。



  控 制 探 針

為確保晶片數據品質,華聯技術團隊經過一連串的測試及驗證,設計一系列的品質控制探針, 以監控完整的晶片實驗步驟,包含樣品製備過程中,miRNA 的螢光標記效率、晶片雜交以及晶片掃描等。
 
 

使用HmiOA文獻 ( 42 )

 PLOS Genetics. PLOS Genetics doi:10.1371/journal.pgen.1005726.
 fMiRNA-192 and miRNA-204 Directly Suppress lncRNA HOTTIP and Interrupt GLS1-Mediated Glutaminolysis in Hepatocellular Carcinoma 
 
  Abstract
Accumulated evidence demonstrated that long non-coding RNAs (lncRNAs) play a pivotal role in tumorigenesis. However, it is still largely unknown how these lncRNAs were regulated by small ncRNAs, such as microRNAs (miRNAs), at the post-transcriptional level. We here use lncRNA HOTTIP as an example to study how miRNAs impact lncRNAs expression and its biological significance in hepatocellular carcinoma (HCC). LncRNA HOTTIP is a vital oncogene in HCC, one of the deadliest cancers worldwide. In the current study, we identified miR-192 and miR-204 as two microRNAs (miRNAs) suppressing HOTTIP expression via the Argonaute 2 (AGO2)-mediated RNA interference (RNAi) pathway in HCC. Interaction between miR-192 or miR-204 and HOTTIP were further confirmed using dual luciferase reporter gene assays. Consistent with this notion, a significant negative correlation between these miRNAs and HOTTIP exists in HCC tissue specimens. Interestingly, the dysregulation of the three ncRNAs was associated with overall survival of HCC patients. In addition, the posttranscriptional silencing of HOTTIP by miR-192, miR-204 or HOTTIP siRNAs could significantly suppress viability of HCC cells. On the contrary, antagonizing endogenous miR-192 or miR-204 led to increased HOTTIP expression and stimulated cell proliferation. In vivo mouse xenograft model also support the tumor suppressor role of both miRNAs. Besides the known targets (multiple 5’ end HOX A genes, i.e. HOXA13), glutaminase (GLS1) was identified as a potential downstream target of the miR-192/-204-HOTTIP axis in HCC. Considering glutaminolysis as a crucial hallmark of cancer cells and significantly inhibited cell viability after silencingGLS1, we speculate that the miR-192/-204-HOTTIP axis may interrupt HCC glutaminolysis through GLS1 inhibition. These results elucidate that the miR-192/-204-HOTTIP axis might be an important molecular pathway during hepatic cell tumorigenesis. Our data in clinical HCC samples highlight miR-192, miR-204 and HOTTIP with prognostic and potentially therapeutic
   

 Prostate. doi: 10.1002/pros.23068. Epub 2015 Aug 26..
 Hsa-miR-146a-5p modulates androgen-independent prostate cancer cells apoptosis by targeting ROCK1
 
 
 
  Abstract
Background MicroRNAs (miRNAs) have been demonstrated playing important roles in the procession of prostate cancer cells transformation from androgen-dependence to androgen-independence. Methods We conducted the miRNA microarray and realtime PCR analyses in both androgen-dependent (ADPC) and androgen-independent prostate cancer (AIPC) tissues. We also explored the role of hsa-miR-146a-5p (miR-146a) in MSKCC prostate cancer clinical database. Moreover, the impact of miR-146a on prostate cancer cells apoptosis were detected by Hoechst staining and fluorescence-activated cell sorter (FACS). Its target is predicted by DIANA LAB online database and the result was assumed by western blotting and luciferase assay. Results We demonstrated that miR-146a was down-regulated in AIPC tissues and cell lines compared to that in the ADPC tissues. In MSKCC data re-analyses, we found that miR-146a was underexpressed in metastatic prostate cancer tissues and those with Gleason score >8, moreover, low level of miR-146a represented a high biochemical relapse rate after radical prostatectomy. In the functional analyses, we transfected miR-146a mimics into CPRC cell lines and found miR-146a induced cells apoptosis. In mechanic analyses, we found that miR-146a inhibited the basal level of Rho-associated, coiled-coil containing protein kinase 1 (ROCK1) expression by targeting its 3'UTR and an inverse correlation of expression between miR-146a and ROCK1 was observed. Moreover, caspase 3 activity was stimulated by miR-146a overexpression. Conclusion miR-146a has a critical role in the process of AIPC prostate cancer cells apoptosis through regulation of ROCK/Caspase 3 pathway. Targeting this pathway may be a promising therapeutic strategy for future personalized anti-cancer treatment.
   

 Nature Cell Biology. 2015, 17(3):311-21. doi: 10.1038/ncb3110.
 Reduced adenosine-to-inosine miR-455-5p editing promotes melanoma growth and metastasis
 
 
 Einav Shoshan, Aaron K. Mobley, Russell R. Braeuer, Takafumi Kamiya, Li Huang, Mayra E. Vasquez, Ahmad Salameh, Ho Jeong Lee, Sun Jin Kim, Cristina Ivan, Guermarie Velazquez-Torres, Ka Ming Nip, Kelsey Zhu, Denise Brooks, Steven J. M. Jones, Inanc Birol,Maribel Mosqueda, Yu-ye Wen, Agda Karina Eterovic, Anil K. Sood, Patrick Hwu, Je rey E. Gershenwald, A. Gordon Robertson, George A. Calin, GalMarkel, Isaiah J. Fidler, Menashe Bar-Eli
  Abstract
Although recent studies have shown that adenosine-to-inosine (A-to-I) RNA editing occurs in microRNAs (miRNAs), its effects on tumour growth and metastasis are not well understood. We present evidence of CREB-mediated low expression of ADAR1 in metastatic melanoma cell lines and tumour specimens. Re-expression of ADAR1 resulted in the suppression of melanoma growth and metastasis in vivo. Consequently, we identified three miRNAs undergoing A-to-I editing in the weakly metastatic melanoma but not in strongly metastatic cell lines. One of these miRNAs, miR-455-5p, has two A-to-I RNA-editing sites. The biological function of edited miR-455-5p is different from that of the unedited form, as it recognizes a different set of genes. Indeed, wild-type miR-455-5p promotes melanoma metastasis through inhibition of the tumour suppressor gene CPEB1. Moreover, wild-type miR-455 enhances melanoma growth and metastasis in vivo, whereas the edited form inhibits these features. These results demonstrate a previously unrecognized role for RNA editing in melanoma progression.
   

 Tumor Biology. 2015, 36(1):219-25. doi: 10.1007/s13277-014-2622-5.
 miR-1285-3p acts as a potential tumor suppressor miRNA via downregulating JUN expression in hepatocellular carcinoma
 
 
 Jibing Liu, Jingchen Yan, Changchun Zhou, Zhenbin Yang, Qinghua Ma, Qingyan Jin
  Abstract
In the world, hepatocellular carcinoma (HCC) is one of the most common and most lethal cancers. Currently, standard therapy for unresectable HCC is a local-regional therapy with transarterial chemoembolisation (TACE). In this study, we sought to assess whether plasma circulating microRNAs (miRNAs) can be used to predict the prognosis of HCC patients receiving the TACE treatment. Firstly, we systematically examined TACE therapeutic effectiveness-related circulating miRNAs through miRNA Profiling Chips. As a result, we identified 19 circulating miRNAs to be significantly differentially expressed between the TACE-response group and the TACE-nonresponse group. In the second stage, we performed quantitative analyses of these candidate miRNAs in additional HCC patients treated with TACE and validated two of the aforementioned 19 miRNAs (miR-1285-3p and miR-4741) as candidate biomarkers for predicting prognosis of TACE. Interestingly, we found that miR-1285-3p could directly repress JUN oncogene expression in HCC cells, indicating miR-1285-3p could act as a potential tumor suppressor. In conclusion, our data indicate that circulating miR-1285-3p and miR-4741 was predictive of response to TACE therapy in HCC.
   

 Cellular physiology and biochemistry. 2015, 35(6):2169-80. doi: 10.1159/000374022.
 MiR-10b Directly Targets ZEB1 and PIK3CA to Curb Adenomyotic Epithelial Cell Invasiveness via Upregulation of E-Cadherin and Inhibition of Akt Phosphorylation
 
 
 Xiao Lang, Zhen Lu, Jianchao Wang, Ting Li, Ying Loao, Chunyan Jia, Wenxia Zhao, Huiqi Fang, Ying Guo
  Abstract
BACKGROUND/AIMS: Adenomyosis is a disease in which ectopic endometrial glands and stromal cells appear in the uterine myometrium. Despite its prevalence, the molecular mechanisms involved in the development of adenomyosis are largely unknown. The aim of this study was to investigate the role of miR-10b and its target genes ZEB1 and PIK3CA in adenomyosis. METHODS: 1387 miRNAs in human normal endometrium and ectopic endometrial lesions of adenomyosis using a microarray screen assay. The significant differential expression of 10 miRNAs was confirmed by qRT-PCR. The expression of miR-10b in endometrial epithelial cells isolated from normal endometrium and paired eutopic and ectopic endometrium of adenomyosis was measured by qRT-PCR. Subsequently, the targets of miR-10b were predicted by bioinformatics and confirmed using a luciferase assay, and the mRNA and protein expression of ZEB1 and PIK3CA were assessed in the endometrium or endometrial epithelial cells by qRT-PCR and western blotting or immunohistochemical analysis. Cell migration and cell invasion of endometrial epithelial cells with different treatments by Transwell assays. The expression of p-AKT, Akt and E-cadherin proteins was determined by Western blot analysis. RESULTS: MiR-10b expression was significantly downregulated in both adenomyotic lesions and adenomyotic epithelial cells. MiR-10b overexpression in adenomyotic epithelial cells inhibited cell migration and invasion. We then demonstrated that miR-10b directly targets the 3'-UTRs of ZEB1 and PIK3CA, and downregulates ZEB1 and PIK3CA in adenomyotic epithelial cells, leading to increased E-cadherin expression and decreased Akt phosphorylation.
   

 BMC Genomics. 2015, 16:501. doi: 10.1186/s12864-015-1642-x.
 Reorganization of metastamiRs in the evolution of metastatic aggressive neuroblastoma cells
 
 
 Faizan H Khan, Vijayabaskar Pandian, Satishkumar Ramraj, Sheeja Aravindan, Terence S Herman, Natarajan Aravindan
  Abstract
Background: MetastamiRs have momentous clinical relevance and have been correlated with disease progression in many tumors. In this study, we identified neuroblastoma metastamiRs exploiting unique mouse models of favorable and high-risk metastatic human neuroblastoma. Further, we related their deregulation to the modulation of target proteins and established their association with clinical outcomes. Results: Whole genome miRNA microarray analysis identified 74 metastamiRs across the manifold of metastatic tumors. RT-qPCR on select miRNAs validated profile expression. Results from bio-informatics across the ingenuity pathway, miRCancer, and literature data-mining endorsed the expression of these miRNAs in multiple tumor systems and showed their role in metastasis, identifying them as metastamiRs. Immunoblotting and TMA-IHC analyses revealed alterations in the expression/phosphorylation of metastamiRs¡¦ targets, including ADAMTS-1, AKT1/2/3, ASK1, AURK£], Birc1, Birc2, Bric5, £]-CATENIN, CASP8, CD54, CDK4, CREB, CTGF, CXCR4, CYCLIN-D1, EGFR, ELK1, ESR1, CFOS, FOSB, FRA, GRB10, GSK3£], IL1£, JUND, kRAS, KRTAP1, MCP1, MEGF10, MMP2, MMP3, MMP9, MMP10, MTA2, MYB, cMYC, NF2, NOS3, P21, pP38, PTPN3, CLEAVED PARP, PKC, SDF-1£], SEMA3D, SELE, STAT3, TLR3, TNF£, TNFR1, and VEGF in aggressive cells ex vivo and in a manifold of metastatic tumors in vivo. miRNA mimic (hsa-miR-125b, hsa-miR-27b, hsa-miR-93, hsa-miR-20a) and inhibitor (hsa-miR-1224-3p, hsa-miR-1260) approach for select miRNAs revealed the direct influence of the altered metastamiRs in the regulation of identified protein targets. Clinical outcome association analysis with the validated metastamiRs¡¦ targets corresponded strongly with poor overall and relapse-free survival. Conclusions: For the first time, these results identified a comprehensive list of neuroblastoma metastamiRs, related their deregulation to altered expression of protein targets, and established their association with poor clinical outcomes. The identified set of distinctive neuroblastoma metastamiRs could serve as potential candidates for diagnostic markers for the switch from favorable to high-risk metastatic disease.
   

 Oncology Reports. 2015, 34(1):318-24. doi: 10.3892/or.2015.3953.
 Microarray analysis of the aberrant microRNA expression pattern in gliomas of different grades
 
 
 Xiao-Peng Zhu, Ke-Jie Mou, Qing-Fu Xu, Jun-Hai Tang, Guo-Hao Huang, Jian-Ping Xu, Guang-Hui Li, Si-Jin Ai, Jean-Phillippe Hugnot, Zheng Zhou, Sheng-Qing Lv
  Abstract
Previous studies have focused on miRNA expression in brain gliomas. However, both the expression pattern of miRNAs in gliomas of different grades and various miRNAs involved in malignant progression of gliomas are poorly understood. In the present study, we used miRNA microarray-based screening to investigate the miRNA expression profile in gliomas, which was further verified by qRT-PCR in selected miRNAs. In total, we found 13 differentially expressed miRNAs between gliomas and their matched surrounding tissues. Among them, 12 miRNAs were upregulated and only one (miR-4489) was downregulated compared with the control. Furthermore, the lower expression level of miR-4489 was confirmed by qRT-PCR in 26 glioma samples. Our microarray result revealed 8, 9 and 15 aberrantly expressed miRNAs in gliomas of World Health Organization (WHO) grade II-IV, respectively. Gene Ontology (GO) and Pathway analysis indicated that target genes of the 13 miRNAs were significantly enriched in central nervous system- and tumor‑related biological processes and signaling pathways. The dysregulated miRNAs identified in the present study contribute to the tumorigenesis and malignant progression of gliomas and may serve as useful markers for advanced glioma pathological grading and prognosis.
   

 Molecular Medicine Reports. 2015, 12(3):3525-30. doi: 10.3892/mmr.2015.3835.
 Identification of a microRNA signature in endothelial cells with mechanical stretch stimulation
 
 
 Jubing Zheng, Kui Zhang, Yueli Wang, Jian Cao, Feng Zhang, Jubing Zheng, Ran Dong
  Abstract
The current study aimed to verify an miRNA signature in endothelial cells undergoing mechanical stretch stimulation. In the present study, microarray profiling was conducted in order to identify the differential expression of miRNAs in endothelial cells undergoing mechanical stimulation, compared with unstimulated endothelial cells. The microarray data was then validated by reverse transcription‑quantitative polymerase chain reaction. Genes and signaling pathways regulated by the miRNAs were investigated in silico using Gene Ontology and the Kyoto Encyclopedia of Genes or Genomes, which are ontological and network‑mapping algorithms. The microarray data collected demonstrated that 38 miRNAs exhibited significant differential expression in endothelial cells with mechanical stretch stimulation. Of these, 20 were upregulated and 18 were downregulated. The results from the in silico analysis indicated that the miRNAs identified were participants in mechanical stretch‑induced endothelial dysfunction. During the initial stage of vein graft failure, which is induced by endothelial dysfunction, a unique miRNA signature was identified. The identified miRNAs are suggested to be involved in the pathological processes of traumatic injury.
   

 Neurobiology of Aging. 2015, 36(3):1356-68. doi: 10.1016/j.neurobiolaging.2014.11.020.
 The CCAAT/enhancer-binding protein delta/miR135a/thrombospondin 1 axis mediates PGE2-induced angiogenesis in Alzheimer's disease
 
 
 Chiung-Yuan Ko, Yu-Yi Chu, Shuh Narumiya, Jhih-Ying Chi, Tomoyuki Furuyashiki, Tomohiro Aoki, Shao-Ming Wang, Wen-Chang Chang, Ju-Ming Wang
  Abstract
In Alzheimer's disease (AD), large populations of endothelial cells undergo angiogenesis due to brain hypoxia and inflammation. Substantial evidence from epidemiologic, pathologic, and clinical reports suggests that vascular factors are critical for the pathogenesis of AD. However, the precise mechanistic correlation between inflammation and angiogenesis in AD has not been well elucidated. Prostaglandin E2 (PGE2), a key factor of the inflammatory response, has been known to promote angiogenesis. In this study, we demonstrated that PGE2 acts through EP4 receptor and protein kinase A to modulate CCAAT/enhancer-binding protein delta (CEBPD) abundance in astrocytes. Attenuated vessel formation was observed in the brains of AppTg/Cebpd(-/-) mice. We showed that miR135a was responsive to the induction of CEBPD and further negatively regulated thrombospondin 1 (THBS1) transcription by directly targeting its 3'-untranslated region (3'UTR) in astrocytes. Furthermore, conditioned media from astrocytes expressing miR135a promoted Human umbilical vein endothelial cells (HUVECs) tube-like formation, which correlated with the effects of PGE2 on angiogenesis. Our results indicated that CEBPD contributes to the repression of THBS1 transcription by activating the expression of miR135a in astrocytes following PGE2 treatment. We provided new evidence that astrocytic CEBPD increases angiogenesis during AD pathogenesis. This discovery supports the negative influence of CEBPD activation in astrocytes with respect to AD pathogenesis and implies that the CEBPD/miR135a/THBS1 axis could be a therapeutic target of AD.
   

 International Journal of Clinical and Experimental Medicine. 2014, 7(12): 5226¡V5234.
 Aberrant expression of microRNAs in serum may identify individuals with pancreatic cancer
 
 
 Wei-Chang Chen, Heng-Jun Gao, Hai-Hui Sheng, Mao-Song Lin, Jun-Xing Huang
  Abstract
Pancreatic cancer (PC) has the poorest survival rate among all types of human cancer due to the lack of sensitive and non-invasive diagnostic screen methods for PC screening. Our aim was to identify novel serum microRNA (miRNA) biomarkers for the early detection of PC. We used microarray to screen differential expression of miRNAs in two pooled serum samples (6 PC patients and 6 healthy controls). A panel of miRNAs (22 over-expression and 23 decreased) were deregulated in serum of PC patients in comparison to controls. The expressions of 8 selected miRNAs were further evaluated in sera from 49 PC patients and 27 controls using quantitative reverse transcription-polymerase chain reaction. The levels of serum miR-492 and miR-663a were significantly decreased in PC patients compared with controls (P < 0.05). ROC curve analysis showed that serum miR-492 and miR-663a yield an AUC of 0.787 with 75.5% sensitivity and 70.0% specificity and 0.870 with 85.7% sensitivity and 80.0% specificity, respectively, for discriminating between PC patients and healthy controls. In addition, the level of miR-663a was significantly and inversely associated with TNM stage (P = 0.027). These results suggested that serum miR-492 and miR-663a could have strong potential as novel non-invasive biomarkers for the early detection of PC.
   

 Oncogene. 2014 Jun 23. doi: 10.1038/onc.2014.177.
 HSF1 regulation of £]-catenin in mammary cancer cells through control of HuR/elavL1 expression
 
 
 SK Calderwood, S-D Chou, A Murshid, T Eguchi, J Gong
  Abstract
There is now compelling evidence to indicate a place for heat shock factor 1 (HSF1) in mammary carcinogenesis, tumour progression and metastasis. Here we have investigated a role for HSF1 in regulating the expression of the stem cell renewal factor £]-catenin in immortalized humanmammary epithelial and carcinoma cells. We found HSF1 to be involved in regulating the translation of £]-catenin, by investigating effects of gain and loss of HSF1 on this protein. Interestingly, although HSF1 is a potent transcription factor, it was not directly involved in regulating levels of £]-catenin mRNA. Instead, our data suggest a complex role in translational regulation. HSF1 was shown to regulate levels of the RNA-binding protein HuR that controlled £]-catenin translation. An extra complexity was added to this scenario when it was shown that the long non-coding RNA molecule lincRNA-p21, known to be involved in £]-catenin mRNA (CTNNB1) translational regulation, was controlled by HSF1 repression. We have shown previously thatHSF1 was positively regulated through phosphorylation by mammalian target of rapamycin (mTOR) kinase on a key residue, serine 326, essential for transcriptional activity. In this study, we found that mTOR knockdown not only decreased HSF1-S326 phosphorylation in mammary cells, but also decreased £]-catenin expression through a mechanism requiring HuR. Our data point to a complex role for HSF1 in the regulation of HuR and £]-catenin expression that may be significant in mammary carcinogenesis.
   

 Scientific Reports. 2014 Oct 6;4:6527. doi: 10.1038/srep06527.
 Reduced miR-3127-5p expression promotes NSCLC proliferation/invasion and contributes to dasatinib sensitivity via the c-Abl/Ras/ERK pathway
 
 
 Bo Su, Wen Gao, Yifeng Sun, Chang Chen, Peng Zhang, Huikang Xie, Likun Hou, Zheng Hui, Yongjie Xu, Qiaoling Du, Xiao Zhou
  Abstract
miR-3127-5p is a primate-specific miRNA which is down-regulated in recurrent NSCLC tissue vs. matched primary tumor tissue (N = 15) and in tumor tissue vs. normal lung tissue (N = 177). Reduced miR-3127-5p expression is associated with a higher Ki-67 proliferation index and unfavorable prognosis in NSCLC. Overexpression of miR-3127-5p significantly reduced NSCLC cells proliferation, migration, and motility in vitro and in vivo. The oncogene ABL1 was a direct miR-3127-5p target, and miR-3127-5p regulated the activation of the Abl/Ras/ERK pathway and transactivated downstream proliferation/metastasis-associated molecules. Overexpression of miR-3127-5p in A549 or H292 cells resulted in enhanced resistance todasatinib, an Abl/src tyrosine kinase inhibitor. miR-3127-5p expression levels were correlated with dasatinib sensitivity in NSCLC cell lines without K-Ras G12 mutation. In conclusion, miR-3127-5p acts as a tumor suppressor gene and is a potential biomarker for dasatinib sensitivity in the non-mutated Ras subset of NSCLC.
   

 Frontiers in Genetics. 2014, 5:246. doi: 10.3389/fgene.2014.00246.
 An investigation into anti-proliferative effects of microRNAs encoded by the miR-106a-363 cluster on human carcinoma cells and keratinocytes using microarray profiling of miRNA transcriptomes
 
 
 Cuong Khuu, Anne-MartheJevnaker, MagneBryne, HaraldOsmundsen
  Abstract
Transfection of human oral squamous carcinoma cells (clone E10) with mimics for unexpressed miR-20b or miR-363-5p, encoded by the miR-106a-363 cluster (miR-20b, miR-106a, miR-363-3p, or miR-363-5p), caused 40-50% decrease in proliferation. Transfection with mimics for miR-18a or miR-92a, encoded by the miR-17-92 cluster (all members being expressed in E10 cells), had no effect on proliferation. In contrast, mimic for the siblingmiRNA-19a yielded about 20% inhibition of proliferation. To investigate miRNA involvement profiling of miRNA transcriptomes were carried out using deoxyoligonucleotide microarrays. In transfectants for miR-19a, or miR-20b or miR-363-5p most differentially expressed miRNAs exhibited decreased expression, including some miRNAs encoded in paralogous miR-17-92-or miR-106b-25 cluster. Only in cells transfected with miR-19a mimic significantly increased expression of miR-20b observed-about 50-fold as judged by qRT-PCR. Further studies using qRT-PCR showed that transfection of E10 cells with mimic for miRNAs encoded by miR-17-92 - or miR-106a-363 - or the miR-106b-25 cluster confirmed selective effect on expression on sibling miRNAs. We conclude that high levels of miRNAs encoded by the miR-106a-363 cluster may contribute to inhibition of proliferation by decreasing expression of several sibling miRNAs encoded by miR-17-92 or by the miR-106b-25 cluster. The inhibition of proliferation observed in miR-19a-mimic transfectants is likely caused by the miR-19a-dependent increase in the levels of miR-20b and miR-106a. Bioinformatic analysis of differentially expressed miRNAs from miR-106a, miR-20b and miR-363-5p transfectants, but not miR-92a transfectants, yielded significant associations to "Cellular Growth and Proliferation" and "Cell Cycle." Western blotting results showed that levels of affected proteins to differ between transfectants, suggesting that different anti-proliferative mechanisms may operate in these transfectants.
   

 Molecular Carcinogenesis. 2014 Sep 22. doi: 10.1002/mc.22221.
 MicroRNA-191, by promoting the EMT and increasing CSC-like properties, is involved in neoplastic and metastatic properties of transformed human bronchial epithelial cells
 
 
 Qizhan Liu, Wenchao Xu, Jie Ji, Yuan Xu, Yawei Liu, Le Shi, Yi Liu, Xiaolin Lu, Yue Zhao, Fei Luo, Bairu Wang, Rongrong Jiang, Jianping Zhang
  Abstract
Lung cancer is the leading cause of cancer mortality worldwide. A common interest in lung cancer research is the identification of biomarkers for early diagnosis and accurate prognosis. There is increasing evidence that microRNAs (miRNAs) are involved in lung cancer. To explore new biomarkers of chemical exposure in risk assessment of chemical carcinogenesis and lung cancer, we analyzed miRNA expression profiles of human bronchialepithelial (HBE) cells malignantly transformed by arsenite. High-throughput microarray analysis showed that 51 miRNAs were differentially expressed in transformed HBE cells relative to normal HBE cells. In particular, miR-191 was up-regulated in transformed cells. In HBE cells, arsenite induced increases of miR-191 and WT1 levels, decreased BASP1 expression, and activated the Wnt/£]-catenin pathway, effects that were blocked by miR-191 knockdown. In addition, a luciferase reporter assay indicated that BASP1 is a direct target of miR-191. By inhibiting the expression of BASP1, miR-191 increased the expression of WT1 to promote activation of Wnt/£]-catenin pathway. In transformed cells, inhibition of miR-191 expression blocked the epithelial-mesenchymal transition (EMT) and cancer stem cell (CSC)-like properties of cells and decreased their migratory capacity andneoplastic properties. Thus, these results demonstrate that miR-191 modulates the EMT and the CSC-like properties of transformed cells and indicate that it is an onco-miR involved in the neoplastic and metastatic properties of transformed cells. 
   

 Tumor Biology. 2014 Sep 18.
 miR-1285-3p acts as a potential tumor suppressor miRNA via downregulating JUN expression in hepatocellular carcinoma
 
 
 Jibing Liu, Jingchen Yan, Changchun Zhou, Qinghua Ma, Qingyan Jin, Zhenbin Yang
  Abstract
In the world, hepatocellular carcinoma (HCC) is one of the most common and most lethal cancers. Currently, standard therapy for unresectable HCC is a local-regional therapy with transarterial chemoembolisation (TACE). In this study, we sought to assess whether plasma circulating microRNAs (miRNAs) can be used to predict the prognosis of HCC patients receiving the TACE treatment. Firstly, we systematically examined TACE therapeutic effectiveness-related circulating miRNAs through miRNA Profiling Chips. As a result, we identified 19 circulating miRNAs to be significantly differentially expressed between the TACE-response group and the TACE-nonresponse group. In the second stage, we performed quantitative analyses of these candidate miRNAs in additional HCC patients treated with TACE and validated two of the aforementioned 19 miRNAs (miR-1285-3pand miR-4741) as candidate biomarkers for predicting prognosis of TACE. Interestingly, we found that miR-1285-3p could directly repress JUNoncogene expression in HCC cells, indicating miR-1285-3p could act as a potential tumor suppressor. In conclusion, our data indicate that circulatingmiR-1285-3p and miR-4741 was predictive of response to TACE therapy in HCC.
   

 BioMed Research International. 2014 Sep 16.
 MicroRNA Expression Profiling Altered by Variant Dosage of Radiation Exposure
 
 
 Kuei-Fang Lee, Yi-Cheng Chen, Paul Wei-Che Hsu, Ingrid Y. Liu, Lawrence Shih-Hsin Wu
  Abstract
Various biological effects are associated with radiation exposure. Irradiated cells may elevate the risk for genetic instability, mutation, and cancer under low levels of radiation exposure, in addition to being able to extend the postradiation side effects in normal tissues. Radiation-induced bystander effect (RIBE) is the focus of rigorous research as it may promote the development of cancer even at low radiation doses. Alterations in the DNA sequence could not explain these biological effects of radiation and it is thought that epigenetics factors may be involved. Indeed, some microRNAs (or miRNAs) have been found to correlate radiation-induced damages and may be potential biomarkers for the various biological effects caused by different levels of radiation exposure. However, the regulatory role that miRNA plays in this aspect remains elusive. In this study, we profiled the expression changes in miRNA under fractionated radiation exposure in human peripheral blood mononuclear cells. By utilizing publicly available microRNA knowledge bases and performing cross validations with our previous gene expression profiling under the same radiation condition, we identified various miRNA-gene interactions specific to different doses of radiation treatment, providing new insights for the molecular underpinnings of radiation injury.
   

 Tumor Biology. 2014 Jul 16.
 MiR-7-5p is frequently downregulated in glioblastoma microvasculature and inhibits vascular endothelial cell proliferation by targeting RAF1
 
 
 Zhiguo Liu, Yuguang Liu, Lianling Li, Zhenkuan Xu, Baibin Bi, Jian Yi Li, Yunyan Wang
  Abstract
The aberrant expression of microRNAs (miRNAs) is always associated with tumor development and progression. Microvascular proliferation is one of the unique pathologic features of glioblastoma (GBM) . In this study, the microvasculature from GBM or normal brain tissue derived from neurosurgeries was purified and total RNA was isolated from purified microvasculature. The difference of miRNA expression profiles betweenglioblastoma microvasculature and normal brain capillaries was investigated. It was found that miR-7-5p in GBM microvessels was significantly reduced compared with that in normal brain capillaries. In the in vitro experiments, overexpression of miR-7-5p significantly inhibited human umbilical vein endothelial cell proliferation. Forced expression of miR-7-5p in human umbilical vein endothelial cells in vitro significantly reduced the protein level of RAF1 and repressed the activity of the luciferase, a reporter vector carrying the 3'-untranslated region of RAF1. These findings indicate that RAF1 is one of the miR-7-5p target genes. Furthermore, a significant inverse correlation between miR-7-5p expression and RAF1 protein level in GBMmicrovasculature was found. These data suggest that miR-7-5p functions as a tumor suppressor gene to regulate GBM microvascular endothelial cellproliferation potentially by targeting the RAF1 oncogene, implicating an important role for miR-7-5p in the pathogenesis of GBM. It may serve as a guide for the antitumor angiogenesis drug development.
   

 BioMed Research International. 2014 July 1.
 Systematic expression profiling analysis identifies specific microRNA-gene interactions that may differentiate between active and latent tuberculosis infection
 
 
 Lawrence Shih-Hsin Wu, Shih-Wei Lee, Kai-Yao Huang, Tzong-Yi Lee, Paul Wei-Che Hsu, Julia Tzu-Ya Weng
  Abstract
Tuberculosis (TB) is the second most common cause of death from infectious diseases. About 90% of those infected are asymptomatic¡Xthe so-called latent TB infections (LTBI), with a 10% lifetime chance of progressing to active TB. In attempt to further understand the molecular pathogenesis of TB and develop efficient diagnostic biomarkers, several molecular studies have attempted to compare the gene and microRNA expression profiles between healthy controls versus active TB or LTBI patients. However, the results vary due to diverse genetic background, study designs, and the inherent complexity of the disease process. Thus, developing a sensitive and efficient method for the detection of LTBI is both crucial and challenging. For the present study, we performed a systematic analysis of the gene and microRNA profiles of healthy individuals versus those affected with TB or LTBI. Combined with a series of in silico analysis utilizing publicly available microRNA knowledge bases and published literature data, we have uncovered several microRNA-gene interactions that specifically target both the blood and lungs, presenting to be useful molecular signatures to help enhance the understanding of TB pathogenesis. Furthermore, some of these molecular interactions are novel, and may serve as potential biomarkers of TB and LTBI, facilitating the development for a more sensitive, efficient, and cost-effective diagnostic assay for TB and LTBI for the Taiwanese population.
   

 BBA Molecular Cell Research. 2014 May 21. doi: 10.1016/j.bbamcr.2014.05.006..
 The protein phosphatase 2A regulatory subunit B55£ is a modulator of signaling and microRNA expression in acute myeloid leukemia cells
 
 
 Vivian R. Ruvolo, Rodrigo Jacamo, JaredK.Burks, Zhihong Zheng, Seshagiri R. Duvvuri, Liran Zhou, Yihua Qiu, Kevin R. Coombes, Nianxiang Zhang, Suk Y. Yoo, Rongqing Pan, NumsenHail Jr., Marina Konopleva, George Calin, Steven M. Kornblau, Michael Andreeff, Peter P. Ruvolo
  Abstract
We recently discovered that the protein phosphatase 2A (PP2A) B55£ subunit (PPP2R2A) is under-expressed in primary blast cells and is unfavorable for remission duration in AML patients. In this study, reverse phase protein analysis (RPPA) of 230 proteins in 511 AML patient samples revealed a strong correlation of B55£ with a number of proteins including MYC, PKC £, and SRC. B55£ suppression in OCI-AML3 cells by shRNA demonstrated that the B subunit is a PKC£ phosphatase. B55£ does not target SRC, but rather the kinase suppresses protein expression of the Bsubunit. Finally, the correlation between B55£ and MYC levels reflected a complex stoichiometric competition between B subunits. Loss of B55£ in OCI-AML3 cells did not change global PP2A activity and the only isoform that is induced is the one containing B56£. In cells containing B55£ shRNA, MYC was suppressed with concomitant induction of the competing B subunit B56£ (PPP2R5A). A recent study determined that FTY-720, a drug whose action involves the activation of PP2A, resulted in the induction of B55£ In AML cells, and a reduction of the B subunit rendered thesecells resistant to FTY-720. Finally, reduction of the B subunit resulted in an increase in the expression of miR-191-5p and a suppression of miR-142-3p. B55£ regulation of these miRs was intriguing as high levels of miR-191 portend poor survival in AML, and miR-142-3p is mutated in 2% of AML patient samples. In summary, the suppression of B55£ activates signaling pathways that could support leukemia cell survival.
   

 Asia Pacific Journal of Clinical Nutrition. 2014, 23(2):331-7. doi: 10.6133/apjcn.2014.23.2.20.
 MicroRNA-125a-3p expression in abdominal adipose tissues is associated with insulin signalling gene expressions in morbid obesity: observations in Taiwanese
 
 
 Chiu-Li Yeh, I-Chi Cheng, Yu-Chen Hou, Weu Wang, Sung-Ling Yeh
  Abstract
Background: Micro (mi) RNAs have been found to play an important role in the regulation of adipogenesis and insulin sensitivity. However, associations between miRNA and insulin signalling-related gene expressions in abdominal adipose tissues in obese subjects remain unclear. Methods: We used a microarray platform to screen miRNA expressions in abdominal adipose tissues between genders in severely obese subjects and found that the top-ranking miRNA in abdominal omental adipose tissues was miRNA-125a-3p. MicroR-125a-3p and insulin signalling-related gene expressions in abdominal omental adipose tissues of all subjects (11 men and 10 women) were subsequently quantified by a real-time PCR. Also, associations of miR-125a-3p with insulin signallingrelated gene expression and biochemical markers in obese subjects were analyzed by a linear regression analysis. Results: miR-125a-3p expressed by abdominal omental adipose tissues was much higher in obese men than women. No gender difference was observed in abdominal subcutaneous adipose tissues. Concomitant with high miR-125a-3p, c-Jun N-terminal kinase gene expression was also higher, whereas insulin receptor was lower in men than women. There were negative associations of miR-125a-3p with the insulin receptor and phosphatidylinositol 3-kinase expressions. Fasting plasma glucose and cholesterol levels were positively associated with miR-125a-3p expression. These associations were obvious in obese men but not women. Conclusion: Our results support the involvement of miR-125a-3p in regulating the insulin signalling pathway and imply that increased miR-125a-3p expression in omental adipose tissues may be a characteristic feature of insulin resistance in obese men.
   

 Journal of Cellular Biochemistry. 2014 Feb 12. doi: 10.1002/jcb.24786.
 Regulatory Roles of miRNA in the Human Neural Stem Cell Transformation to Glioma Stem Cells
 
 
 Shuang Liu, Jianning Zhang, Max S. Wicha, Alfred E. Chang, Wenhong Fan, Ling Chen, Ming Fan, Qiao Li, Feng Yin
  Abstract
To investigate the expressional alternation of microRNAs (miRNA) during the malignant transformation and development of human glioma, we measuredmiRNA expression profile as well as mRNA expression profile in normal human neural stem cells (hNSCs) and human glioma stem cells (hGSCs). We found 116 miRNA up-regulated and 62 miRNA down-regulated in GSCs. On the other hand, we identified 1,372 mRNA down-regulated, and 1,501 mRNA up-regulated in GSCs compared to those in NSCs. We then analyzed the pathways and the predicted target genes of the miRNAs which differ significantly in expression between GSCs and NSCs using the statistical enrichment methods. These target mRNAs are involved in many cancer-related signaling pathways, such as cell cycle, axon guidance, glioma development, adhesion junction, MAPK and Wnt signaling. Furthermore, we obtained the differently expressed miRNA-target relationships according to the £c value which is used to calculate the regulation extent of miRNA-target and using the databases of miRanda, Targetscans and Pictar. Among the top 10 miRNA-target relationships, hsa-miR-198 and its potential targeted gene DCX and NNAT were selected for validation, and NNAT was found to be the direct target of miR-198. Finally, the functional roles of miR-155-5p and miR-124-3p whose expressions altered significantly between GSCs and NSCs were addressed. Our results provide new clues for the potential mechanisms involved in the origin and development of glioma. Clinically, the altered miRNAs may serve as potential targets and diagnostic tools for novel therapeutic strategies of glioblastoma.
   

 Biochemical and Biophysical Research Communications. 2014 Feb 28. doi: 10.1016/j.bbrc.2014.02.073.
 miR-138-5p reverses gefitinib resistance in non-small cell lung cancer cells via negatively regulating G protein-coupled receptor 124
 
 
 Yi Gao, XiaoWu Fan, WeiNa Li, Wei Ping, Yu Deng, XiangNing Fu
  Abstract
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR TKIs) such as gefitinib are clinically effective treatments for non-small cell lung cancer(NSCLC) patients with EGFR activating mutations. However, therapeutic effect is ultimately limited by the development of acquired TKI resistance. MicroRNAs (miRNAs) represent a category of small non-coding RNAs commonly deregulated in human malignancies. The aim of this study was to investigate the role of miRNAs in gefitinib resistance. We established a gefitinib-resistant cell model (PC9GR) by continually exposing PC9 NSCLC cells togefitinib for 6months. MiRNA microarray screening revealed miR-138-5p showed the greatest downregulation in PC9GR cells. Re-expression of miR-138-5pwas sufficient to sensitize PC9GR cells and another gefitinib-resistant NSCLC cell line, H1975, to gefitinib. Bioinformatics analysis and luciferase reporter assay showed that G protein-coupled receptor124 (GPR124) was a direct target of miR-138-5p. Experimental validation demonstrated that expression of GPR124 was suppressed by miR-138-5p on protein and mRNA levels in NSCLC cells. Furthermore, we observed an inverse correlation between the expression of miR-138-5p and GPR124 in lung adenocarcinoma specimens. Knockdown of GPR124 mimicked the effects of miR-138-5p on the sensitivity to gefitinib. Collectively, our results suggest that downregulation of miR-138-5p contributes to gefitinib resistance and that restoration of miR-138-5p or inhibition GPR124 might serve as potential therapeutic approach for overcoming NSCLC gefitinib resistance.
   

 Nature Cell Biology. 2014 Feb 23;16(3):268-280. doi: 10.1038/ncb2910.
 MicroRNA-146a directs the symmetric division of Snail-dominant colorectal cancer stem cells
 
 
 Wei-Lun Hwang, Jeng-Kae Jiang, Shung-Haur Yang, Tse-Shun Huang, Hsin-Yi Lan, Hao-Wei Teng, Chih-Yung Yang, Ya-Ping Tsai, Chi-Hung Lin, Hsei-WeiWang, Muh-Hwa Yang
  Abstract
Asymmetrical cell division (ACD) maintains the proper number of stem cells to ensure self-renewal. In cancer cells, the deregulation of ACD disrupts the homeostasis of the stem cell pool and promotes tumour growth. However, this mechanism is unclear. Here, we show a reduction of ACD in spheroid-derived colorectal cancer stem cells (CRCSCs) compared with differentiated cancer cells. The epithelial-mesenchymal transition (EMT) inducer Snail is responsible for the ACD-to-symmetrical cell division (SCD) switch in CRCSCs. Mechanistically, Snail induces the expression of microRNA-146a (miR-146a) through the £]-catenin-TCF4 complex. miR-146a targets Numb to stabilize £]-catenin, which forms a feedback circuit to maintain Wnt activity and directs SCD. Interference with the Snail-miR-146a-£]-catenin loop by inhibiting the MEK or Wnt activity reduces the symmetrical division of CRCSCs and attenuates tumorigenicity. In colorectal cancer patients, the SnailHighNumbLow profile is correlated with cetuximab resistance and a poorer prognosis. This study elucidates a unique mechanism of EMT-induced CRCSC expansion.
   

 International Journal of Molecular Medicine. 2013, 32(3):557-67. doi: 10.3892/ijmm.2013.1424.
 Serum microRNA expression levels can predict lymph node metastasis in patients with early-stage cervical squamous cell carcinoma
 
 
 DESHENG YAO, JUNYING CHEN, YUE LI, HONG CHEN, CHANJUAN HE, NAN DING, YAN LU, TINGYU OU, SHAN ZHAO, LI LI, FENGYI LONG
  Abstract
Circulating microRNA expression levels can serve as diagnostic/prognostic biomarkers in several types of malignant tumors; however, to our knowledge, there have been reports describing their value in cervical squamous cell carcinoma (SCC). In this study, we used hybridization arrays to compare the microRNA expression profiles in cervical squamous cell carcinomas (SCC) samples among patients with lymph node metastasis (LNM) or without LNM; 89 microRNAs were found to fit our inclusion criteria. Using quantitative PCR (qPCR), we examined the expression levels of these microRNAs in cervical cancer tissue, as well as in serum from patients and healthy women. We compared the expression levels between patients with LNM (n=40) and those without LNM (n=40) and healthy controls (n=20). Using regression analysis, we generated a comprehensive set of marker microRNAs and drew the fitted binormal receiver operating characteristic (ROC) curves to access the predictive value. We identified 6 serum microRNAs that can predict LNM in cervical SCC patients; these microRNAs were miR-1246, miR-20a, miR-2392, miR-3147, miR-3162-5p and miR-4484. The area under the curve (AUC) of the comprehensive set of serum microRNAs predicting LNM was 0.932 (sensitivity, 0.856; specificity, 0.850). The predictive value of the serum microRNAs was inferior to that in tissue (AUC 0.992; sensitivity, 0.967; specificity, 0.950; P=0.018). We compared the LNM predictive value of serum microRNAs and SCC antigen (SCC-Ag) by drawing fitted binormal ROC curves However, serum microRNA analysis is by far superior to serum SCC‑Ag analysis (AUC 0.713; sensitivity, 0.612; specificity, 0.700; P<0.0001). Serum microRNAs are a good predictor of LNM with clinical value in early-stage cervical SCC.
   

 European Journal of Neuroscience. 2013 Dec 5. doi: 10.1111/ejn.12444.
 Widespread microRNA dysregulation in multiple system atrophy ¡V disease-related alteration in miR-96
 
 
 Kiren Ubhi, Edward Rockenstein, Christine Kragh, Chandra Inglis, Brian Spencer, Sarah Michael, Michael Mante, Anthony Adame, Douglas Galasko, Eliezer Masliah
  Abstract
MicroRNA (miRNA) are short sequences of RNA that function as post-transcriptional regulators by binding to target mRNA transcripts resulting in translational repression. A number of recent studies have identified miRNA as being involved in neurodegenerative disorders including Alzheimer's disease, Parkinson's disease and Huntington's disease. However, the role of miRNA in multiple system atrophy (MSA), a progressive neurodegenerative disorder characterized by oligodendroglial accumulation of alpha-synuclein remains unexamined. In this context, this study examined miRNA profiles in MSA cases compared with controls and in transgenic (tg) models of MSA compared with non-tg mice. The results demonstrate a widespread dysregulation of miRNA in MSA cases, which is recapitulated in the murine models. The study employed a cross-disease, cross-species approach to identify miRNA that were either specifically dysregulated in MSA or were commonly dysregulated in neurodegenerative conditions such as Alzheimer's disease, dementia with Lewy bodies, progressive supranuclear palsy and corticobasal degeneration or the tg mouse model equivalents of these disorders. Using this approach we identified a number of miRNA that were commonly dysregulated between disorders and those that were disease-specific. Moreover, we identified miR-96 as being up-regulated in MSA. Consistent with the up-regulation of miR-96, mRNA and protein levels of members of the solute carrier protein family SLC1A1 and SLC6A6, miR-96 target genes, were down-regulated in MSA cases and a tg model of MSA. These results suggest that miR-96 dysregulation may play a role in MSA and its target genes may be involved in the pathogenesis of MSA.
   

 Oncology Reports. 2013 Nov 28. doi: 10.3892/or.2013.2877.
 Bioinformatic analysis of the membrane cofactor protein CD46 and microRNA expression in hepatocellular carcinoma
 
 
 ZEJUN LU, CHUANFU ZHANG, JIAJUN CUI, QI SONG, LIGUI WANG, JINGBO KANG, PENG LI, XIAOFENG HU, HONGBIN SONG, JINLIANG YANG, YANSONG SUN
  Abstract
The therapeutic potential of membrane complement regulatory protein (mCRP)-neutralizing antibodies is unsatisfactory, which perhaps lies in the complex role of mCRPs in tumor occurrence and development. As a member of the mCRPs, CD46 is a transmembrane protein with a cytoplasmic domain and is implicated more in the control of the alternative complement pathway than of the classical complement pathway. Growing evidence has revealed that both the CD46 signaling pathway and microRNAs (miRNAs) play an important role in the development and progression of hepatocellular carcinoma (HCC). In the present study, we analyzed mCRP expression in different tumor tissues by employing western blotting and qPCR. To address the potential role of miRNAs in CD46 signaling, we set out to profile miRNA expression in CD46-overexpressed and -silenced HepG2 cell lines. Furthermore, bioinformatic analysis was performed to identify downstream targets of CD46 signaling. We found that the levels of CD46 expression in HCC tissues were significantly higher compared to that in the adjacent normal tissues. After complement-related gene expression profiling and unsupervised hierarchical clustering analysis of 10 HCC tissues, a total of 37 miRNAs showed significantly different expression levels before and after CD46 expression change. By bioinformatic analysis, we identified let-7b and miR-17 as downstream targets of CD46 signaling, and that the expression levels of let-7b and miR-17 were negatively correlated with that of CD46 in HepG2 cells. The present study suggests that CD46 plays an important role in HCC carcinogenesis by regulating let-7b and miR-17.
   

 Asian Journal of Andrology. 2013 Aug 26. doi: 10.1038/aja.2013.80.
 miR-205 is frequently downregulated in prostate cancer and acts as a tumor suppressor by inhibiting tumor growth
 
 
 Wang N, Li Q, Feng NH, Cheng G, Guan ZL, Wang Y, Qin C, Yin CJ, Hua LX
  Abstract
The purpose of this study was to elucidate the molecular mechanisms of microRNA-205 (miR-205) as a tumor suppressor in prostate cancer (PCa). In the present study, microRNA microarray analysis suggested that the expression of miR-205 was significantly decreased in advanced PCa compared with early PCa. Real-time PCR analysis also indicated that miR-205 expression was significantly decreased in PCa tissues compared with non-cancerous tissues. Moreover, the expression of miR-205 has been demonstrated to be associated with the clinicopathological stage and total/free prostate-specific antigen (PSA) level of PCa. Functional analyses showed that both the overexpression of miR-205 and the knockdown of c-SRC in PCa cell lines could inhibit cell growth, colony formation, migration, invasion and the cell cycle as well as induce cell apoptosis in vitro. Furthermore, over-expressing miR-205 reduced tumorigenicity in vivo. Through a luciferase activity assay and Western blotting, c-SRC was identified as a target of miR-205 in cells. The overexpression of miR-205 suppressed c-SRC and its downstream signaling molecules, including FAK, p-FAK, ERK1/2 and p-ERK1/2, and attenuated cell proliferation, invasion and tumor growth.
   

 Molecular Carcinogenesis. 2013 Jul 17. doi: 10.1002/mc.22064.
 Inhibitions of Epithelial to Mesenchymal Transition and Cancer Stem Cells‐Like Properties Are Involved in miR‐148a‐Mediated Anti‐Metastasis of Hepatocellular Carcinoma
 
 
 Han Yan, Xiaogang Dong, Xiaoqin Zhong, Jing Ye, Yun Zhou, Xiaojun Yang, Jian Shen, Jianping Zhang
  Abstract
The epithelial¡Vmesenchymal transition (EMT) and acquisition of cancer stem cells (CSCs)-like properties are essential steps in the metastasis and postsurgical recurrence of hepatocellular carcinomas (HCCs). The molecular mechanisms involved, however, remain obscure. As determined by an miRNA microarray analysis, there was lower expression of miR-148a in poorly differentiated HCC tissues relative to well-differentiated HCC tissues. MHCC97H and MHCC97L (HCC cells with migratory capacity) and HCC tissues with various differentiation status were selected for further investigation. The results showed that miR-148a levels inversely correlated with the differentiation status of HCC tissues. In MHCC97H and MHCC97L cells, over-expression of miR-148a blocked the EMT process, attenuated the expression of CD90 and CD44 (biomarkers for liver cancer stem cells), and inhibited their migratory capacity. Via TargetScan and microRNA.org algorithms, miR-148a was predicted to bind to the Wnt1 mRNA 3'-UTR. Wnt1 was confirmed as a target gene of miR-148a in HCC cells, and the Wnt signal pathway was determined to be involved in the miR-148a-mediated inhibition of EMT and CSCs-like properties of MHCC97H cells. Moreover, the expression of miR-148a in nonmetastatic HCC tissues was higher than that in metastatic HCC tissues. The results suggest that miR-148a inhibits the metastasis of HCCs by blocking EMT and CSCs-like properties through effects on the Wnt signaling pathway.
   

 Cell Reports. 2013, 3(6):2100-12. doi: 10.1016/j.celrep.2013.05.038.
 DNA-Damage-Induced Nuclear Export of Precursor MicroRNAs Is Regulated by the ATM-AKT Pathway
 
 
 Guohui Wan, Xinna Zhang, Robert R. Langley, Yunhua Liu, Xiaoxiao Hu, Cecil Han, Guang Peng, Lee M. Ellis, Stephen N. Jones, Xiongbin Lu
  Abstract
Expression of microRNAs (miRNAs) involves transcription of miRNA genes and maturation of the primary transcripts. Recent studies have shown that posttranscriptional processing of primary and precursor miRNAs is induced after DNA damage through regulatory RNA-binding proteins in the Drosha and Dicer complexes, such as DDX5 and KSRP. However, little is known about the regulation of nuclear export of pre-miRNAs in the DNA-damage response, a critical step in miRNA maturation. Here, we show that nuclear export of pre-miRNAs is accelerated after DNA damage in an ATM-dependent manner. The ATM-activated AKT kinase phosphorylates Nup153, a key component of the nucleopore, leading to enhanced interaction between Nup153 and Exportin-5 (XPO5) and increased nuclear export of pre-miRNAs. These findings define an important role of DNA-damage signaling in miRNA transport and maturation.
   

 Liver International. 2013 Apr 14.
 MicroRNA-491 is Involved in Metastasis of Hepatocellular Carcinoma by Inhibitions of Matrix Metalloproteinase and Epithelial to Mesenchymal Transition
 
 
 Yun Zhou, Yuan Li, Jing Ye, Rongrong Jiang, Han Yan, Xiaojun Yang, Qizhan Liu b, Jianping Zhang
  Abstract
Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related mortality worldwide. The prognosis of HCC patient remains poor due to intrahepatic and extrahepatic metastasis and post-surgical recurrence, however, the mechanisms underlying metastasis and recurrence remain obscure. Here, by employing an miRNAs microarray analysis, we found that miR-491 level was one of the most significant down-regulation in poorly differentiated HCC tissue compared to well differentiated HCC tissue. We then selected HepG2 (very low migratory capacity), MHCC97L (low migratory capacity), and MHCC97H (high migratory capacity) as well as HCC tissues with different status to further investigate the effects of miR-491 on the metastasis of HCC. Our data showed that miR-491 levels were inversely correlated with different status of differentiation in HCC tissues and with migratory potential in HCC cell lines. In HepG2 cells, inhibition of miR-491 increased the expression of matrix metalloproteinase 2/9 (MMP-2/9) and the migratory potential; however, in MHCC97H cells, overexpression of miR-491 level decreased the expression of MMP-2/9 and the migratory capacity. Moreover, miR-491 had a positive relationship with E-cadherin level; however, it had a negative relationship with vimentin level both in cell lines and tissue samples of HCC. MiR-491 levels of non-metastasis HCC tissue are higher than that of metastasis HCC tissue. Our results suggest that miR-491 is involved in metastasis of HCC by blocking EMT and decreasing MMP-9 levels, which may provide a new clue for preventing tumor metastasis of HCC.
   

 Cancer Research. 2013 May 1.
 miR-124 inhibits STAT3 signaling to enhance T cell-mediated immune clearance of glioma
 
 
 Jun Wei, Fei Wang, Ling-Yuan Kong, Shuo Xu, Tiffany Doucette, Sherise D. Ferguson, Yuhui Yang, Kayla McEnery, Krishan Jethwa, Olsi Gjyshi, Wei Qiao, Nicholas B. Levine, Frederick F. Lang, Ganesh Rao, Gregory N. Fuller, George A. Calin, Amy B. Heimberger
  Abstract
MicroRNAs (miRs) have been shown to modulate critical gene transcripts involved in tumorigenesis, but their role in tumor-mediated immune suppression is largely unknown. On the basis of miRNA gene expression in gliomas using tissue microarrays, in situ hybridization, and molecular modeling, miR-124 was identified as a lead candidate for modulating signal transducer and activator of transcription 3 (STAT3) signaling, a key pathway mediating immune suppression in the tumor microenvironment. miR-124 is absent in all grades and pathological types of gliomas. Upon up regulating miR-124 in glioma cancer stem cells (gCSCs), the STAT3 pathway was inhibited, and miR-124 reversed gCSC-mediated immune suppression of T-cell proliferation and induction of Foxp3+ regulatory T-cells (Tregs). Treatment of T-cells from immunosuppressed glioblastoma patients with miR-124 induced marked effector response including up regulation of IL-2, IFN-£^, and tumor necrosis factor (TNF)-£. Both systemic administration of miR-124 or adoptive miR-124-transfected T-cell transfers exerted potent antiglioma therapeutic effects in clonotypic and genetically engineered murine models of glioblastoma and enhanced effector responses in the local tumor microenvironment. These therapeutic effects were ablated in both CD4+ and CD8+ depleted mice and nude mouse systems, indicating that the therapeutic effect of miR-124 depends on the presence of a T-cellmediated antitumor immune response. Our findings highlight the potential application of miR- 124 as a novel immunotherapeutic agent for neoplasms and serve as a model for identifying miRNAs that can be exploited as immune therapeutics.
   

 Evidence-Based Complementary and Alternative Medicine. 2013 March 29.
 A Systems Biology Approach to Characterize Biomarkers for Blood Stasis Syndrome of Unstable Angina Patients by Integrating MicroRNA and Messenger RNA Expression Profiling
 
 
 Jie Wang, Gui Yu
  Abstract
Blood stasis syndrome (BSS) in Traditional Chinese medicine (TCM) was considered to the major type of syndrome in unstable angina (UA) patients, which was proven by the epidemiological investigation. This paper identified the systems biology-based microRNA (miRNA) and mRNA expression biomarkers for BSS of UA. The aim of this study was to compare miRNAs and mRNAs profiles of peripheral blood mononuclear cells (PBMCs) from BSS of UA patients and healthy controls through a systems biology approach. We identified 1081 mRNAs and 25 miRNAs differentially expressed between BSS of UA patients and healthy controls by microarrays. We used DAVID, miRTrail and the protein-protein interactions (PPI) method to explore the related pathways and networks of differentially expressed miRNAs and mRNAs. By combining the results of pathways and networks, we found that the upregulation of miR-146b-5p may induce the downregulation of CALR to attenuate inflammation and the upregulation of miR-199a-5p may induce the downregulation of TP53 to inhibit apoptosis in BSS of UA patients. The expression patterns of miR-146b-5p, miR-199a-5p, CALR and TP53 were confirmed by real-time quantitative polymerase chain reaction (qRT-PCR) in an independent validation cohort including BBS of UA, non-BBS of UA and healthy control. miR-146b-5p, miR-199a-5p, CALR and TP53 could be the biomarkers of BSS of UA patients. The systems biology-based miRNA and mRNA expression biomarkers for the BSS of UA may be helpful for the further stratification of UA patients when deciding on interventions or clinical trials.
   

 PLoS One. 2013, 8(3): e58929. doi:10.1371/journal.pone.0058929.
 Genistein Up-Regulates Tumor Suppressor MicroRNA-574-3p in Prostate Cancer
 
 
 Takeshi Chiyomaru, Soichiro Yamamura, Shinichiro Fukuhara, Hideo Hidaka, Shahana Majid, Sharanjot Saini, Sumit Arora, Guoren Deng, Varahram Shahryari, Inik Chang, Yuichiro Tanaka, Z., Rajvir Dahiya
  Abstract
Genistein has been shown to inhibit cancers both in vitro and in vivo, by altering the expression of several microRNAs (miRNAs). In this study, we focused on tumor suppressor miRNAs regulated by genistein and investigated their function in prostate cancer (PCa) and target pathways. Using miRNA microarray analysis and real-time RT-PCR we observed that miR- 574-3p was significantly up-regulated in PCa cells treated with genistein compared with vehicle control. The expression of miR-574-3p was significantly lower in PCa cell lines and clinical PCa tissues compared with normal prostate cells (RWPE-1) and adjacent normal tissues. Low expression level of miR-574-3p was correlated with advanced tumor stage and higher Gleason score in PCa specimens. Re-expression of miR-574-3p in PCa cells significantly inhibited cell proliferation, migration and invasion in vitro and in vivo. miR-574-3p restoration induced apoptosis through reducing Bcl-xL and activating caspase- 9 and caspase-3. Using GeneCodis software analysis, several pathways affected by miR-574-3p were identified, such as ¡¥Pathways in cancer¡¦, ¡¥Jak-STAT signaling pathway¡¦, and ¡¥Wnt signaling pathway¡¦. Luciferase reporter assays demonstrated that miR-574-3p directly binds to the 39 UTR of several target genes (such as RAC1, EGFR and EP300) that are components of ¡¥Pathways in cancer¡¦. Quantitative real-time PCR and Western analysis showed that the mRNA and protein expression levels of the three target genes in PCa cells were markedly down-regulated with miR-574-3p. Loss-of-function studies demonstrated that the three target genes significantly affect cell proliferation, migration and invasion in PCa cell lines. Our results show that genistein up-regulates tumor suppressor miR-574-3p expression targeting several cell signaling pathways. These findings enhance understanding of how genistein regulates with miRNA in Pca.
   

 Atherosclerosis. 2013 Mar 15. doi: 10.1016/j.atherosclerosis.2013.01.036 .
 A functional polymorphism of PON1 interferes with microRNA binding to increase the risk of ischemic stroke and carotid atherosclerosis
 
 
 Mu-En Liu, Yi-Chu Liao, Ruey-Tay Lin, Yung-Song Wang, Edward His, Hsiu-Fen Lin, Ku-Chung Chen, Suh-Hang Hank Juo
  Abstract
Objective: Single nucleotide polymorphisms (SNPs) located at microRNA (miRNA) binding sites (miRSNPs) can affect the expression of genes. This study aimed to identify the miR-SNPs associated with atherosclerosis and stroke.

Methods: Patients with ischemic stroke (n =657) and stroke- and myocardial infarction-free volunteers (n =1571) were enrolled. The carotid intima-media thickness (IMT) was measured in the control participants. Seventy-nine stroke susceptibility genes were initially selected and 13 genes were predicted to have miR-SNPs at their 3'untranslated regions (3¡¦UTR). The miRNA arrays were used to further identify potential miR-SNPs. The miR-SNP rs3735590 at the paraoxonase 1 (PON1) gene was finally selected and its associations with stroke and carotid IMT were evaluated. The 3¡¦UTR reporter and SNP functional assays were then performed to validate the results.

Results: Compared with CC genotype, patients with CT or TT genotype at rs3735590 had lower risk of ischemic stroke (OR =0.72, p =0.036; OR =0.83, p =0.077, respectively). Among the healthy participants, the CT or TT genotype was associated with thinner IMT in the internal carotid arteries in comparison with CC genotype (£]=-0.76, p =0.003; £]=- 0.022, p =0.452, respectively). Our findings suggested that the minor allele T had a protective effect on atherosclerosis. Results from 3¡¦UTR reporter assays showed that PON1 is a direct target gene of miR-616. In plasmid constructs carrying the risk allele C at rs3735590, miR-616 inhibited the genetic expression of PON1. However, substitution of C by T at rs3735590 reduced the miR-616 binding affinity, leading to overexpression of the PON1 gene.

Conclusion: Our study is the first to show that the miR-SNP at PON1 could affect genetic expression and is associated with an elevated risk for ischemic stroke and subclinical atherosclerosis.
   

 Biochimica et Biophysica Acta. 2013 Feb 8. doi: 10.1016/j.bbagrm.2013.01.011.
 Transfection of siRNAs can alter miRNA levels and trigger non-specific protein degradation in mammalian cells
 
 
 Christopher E. Hart, Stanley T. Crooke, Xue-hai Liang
  Abstract
Sequence-non-specific effects of siRNAs that alter the expression of non-targeted genes have been reported, including competition of siRNAs with endogenous RISC components. However, the detailed mechanisms and subsequent effects of such competition are not well documented. Here we analyze the competition of miRNAs in mammalian cells with low concentrations of siRNAs, and found that: 1) transfection of different siRNAs in the low nanomolar range used to deplete target RNAs can reduce the levels of miRNAs in different cell types, 2) siRNA transfection results in rapid reduction of Ago2-associated miRNAs concurrent with accumulation of Ago2-bound siRNAs and a significant change in the expression levels of many miRNAs, 3) competition largely depends on Ago2 and not Dicer, 4) microarray analysis showed that the majority of highly expressed miRNAs are reduced, in a siRNA concentration dependent manner, and low abundant miRNAs may be unchanged or repressed and a fewmiRNAs appear to have increased levels, and 5) consistent with previous studies, the expres-sion levels ofmRNAs that are targeted by highly repressedmiRNAs are preferentially increased. As a consequence of such competition, we observed that £-tubulin, a substrate of two up-regulated proteases, granzyme B and granzyme M, was rapidly degraded at the protein level upon siRNA transfection. Our results support a model in which transfection of siRNAs can change the levels of many miRNAs by competition for Ago2, leading to altered expression of many miRNA target genes, which can in turn affect downstream gene expression even at the protein level.
   

 Oncology Reports. 2012, 28(6):2115-24. doi: 10.3892/or.2012.2054.
 miRNA expression profile of colon cancer stem cells compared to non-stem cells using the SW1116 cell line
 
 
 ZONGYOU CHEN, YANTIAN FANG, JIANBIN XIANG, XIAODONG GU, ZHENGYANG LI, FENG TANG, ZHONGWEN ZHOU
  Abstract
Colorectal cancer (CRC) is one of the major causes of cancer-related mortality worldwide. Recent studies revealed that there is a relationship between CRC occurrence and microRNA (miRNA) function. Stem cells are a type of cells that have the ability to self-renew and to proliferate extensively while maintaining the undifferentiated state. Cancer stem cells (CSCs) are closely linked to tumor recurrence and metastasis. To this end, we evaluated themiRNA expression differences between colon CSCs and non-stem cells using the SW1116 cell line, to determine the relationship between tumorstem cells and tumor biological behavior. We isolated populations of colon CSCs with the CD133+/CD44+ and CD133-/CD44- surface phenotype from a human SW1116 colon adenocarcinoma cell line using flow cytometry. The expression of miRNA and mRNA of both sets of cells was examined withmiRNA and mRNA arrays. Bioinformatic methods were used to analyze microarray results. We completed gene ontology analysis, pathway analysis,miRNA target gene prediction with databases. We identified a colon stem cell miRNA expression profile comprising 31 upregulated and 31 downregulated miRNAs, such as miR29a, miR29b, miR449b and miR4524. Some of these differentially expressed miRNAs may be involved in the regulation of stem cell differentiation. Gene ontology and pathway analyses showed that the differences are closely related to the function of the cellcycle, cell differentiation, signaling pathway, cytoskeletal proteins and cell-matrix adhesion in colon cancer stem cells. We found that miRNAs play an important role in regulating the expression of colon CSC characteristics. By regulating the expression of CSC signaling pathways, cytoskeleton and membrane proteins, miRNAs give tumor stem cells the macrobiological behavior of recurrence and metastasis. This study provides a new perspective on CRC metastasis and recurrence.
   

 Molecular Cancer Therapeutics. 2012, 11(1):244-53. doi: 10.1158/1535-7163.MCT-11-0592.
 Tumor Suppressor MicroRNA-493 Decreases Cell Motility and Migration Ability in Human Bladder Cancer Cells by Downregulating RhoC and FZD4
 
 
 Koji Ueno, Hiroshi Hirata, Shahana Majid, Soichiro Yamamura, Varahram Shahryari, Z. Laura Tabatabai, Yuji Hinoda, Rajvir Dahiya
  Abstract
The purpose of this study was to identify new tumor suppressor microRNAs (miRNA; miR) in bladder cancer, conduct functional analysis of their suppressive role, and identify their specific target genes. To explore tumor suppressor miRs in bladder cancer, miR microarray was conducted using SV-HUC-1, T24, J82, and TCCSUP cells. Expression of miR-493 in bladder cancer (T24, J82, and TCCSUP) cells was downregulated compared with normal SV-HUC-1cells. Also, the expression of miR-493 was significantly lower in bladder cancer tissues than in their corresponding noncancerous tissues. Transfection of miR-493 into T24 or J82 cells decreased their cell growth and migration abilities. On the basis of this result, to identify potential miR-493 target genes, we used target scan algorithms to identify target oncogenes related to invasion and migration. miR-493 decreased 3'-untranslated region luciferase activity and protein expression of FZD4 and RhoC. miR-493 also decreased binding of RhoC and Rock-1. miR-493 is a new tumor suppressor miRNA in bladder cancer and inhibits cell motility through downregulation of RhoC and FZD4.
   

 PLoS ONE. 2012, 7(1):e30635. doi: 10.1371/journal.pone.0030635 .
 microRNA-152 Mediates DNMT1-Regulated DNA Methylation in the Estrogen Receptor £ Gene
 
 
 Yung-Song Wang, Wen-Wen Chou, Ku-Chung Chen, Hsin-Yun Cheng, Ruey-Tay Lin, Suh-Hang Hank Juo
  Abstract
"Estrogen receptor a (ERa) has been shown to protect against atherosclerosis. Methylation of the ERa gene can reduce ERa expression leading to a higher risk for cardiovascular disease. Recently, microRNAs have been found to regulate DNA methyltransferases (DNMTs) and thus control methylation status in several genes. We first searched for microRNAs involved in DNMT-associated DNA methylation in the ERa gene. We also tested whether statin and a traditional Chinese medicine (San-Huang-Xie-Xin-Tang, SHXXT) could exert a therapeutic effect on microRNA, DNMT and ERa methylation. The ERa expression was decreased and ERa methylation was increased in LPS-treated human aortic smooth muscle cells (HASMCs) and the aorta from rats under a high-fat diet. microRNA-152 was found to be down regulated in the LPS-treated HASMCs. We validated that microRNA-152 can knock down DNMT1 in HASMCs leading to hypermethylation of the ERa gene. Statin had no effect on microRNA-152, DNMT1 or ERa expression. On the contrary, SHXXT could restore microRNA-152, decrease DNMT1 and increase ERa expression in both cellular and animal studies. The present study showed that microRNA-152 decreases under the pro-atherosclerotic conditions. The reduced microRNA-152 can lose an inhibitory effect on DNA methyltransferase, which leads to hypermethylation of the ERa gene and a decrease of ERa level. Although statin can not reverse these cascade proatherosclerotic changes, the SHXXT shows a promising effect to inhibit this unwanted signaling pathway."
   

 Cancer Letters. 2012, 314(2):155-65. doi: 10.1016/j.canlet.2011.09.027.
 Estrogen receptor-regulated microRNAs contribute to the BCL2/BAX imbalance in endometrial adenocarcinoma and precancerous lesions
 
 
 Xueli Zhang, Baolin Xing, Youhua Sheng, Huan Lu, Zhenhong Wei, Rong Zhang, Yifeng He
  Abstract
Uncontrolled estrogen exposure can induce an imbalance in BCL2/BAX expression in endometrial cells, leading to precancerous lesions and type I endometrial adenocarcinoma. This study aimed to explore the mechanism underlying this phenomenon. We show that the activated estrogen receptor can suppress the expression of BAX by upregulating a group of microRNAs including hsa-let-7 family members and hsa-miR-27a, thereby promoting an increased BCL2/BAX ratio as well as enhanced survival and proliferation in the affected cells. These ER-regulated hsa-let-7 microRNAs can be detected in most hyperplastic endometria, suggesting their potential utility as indicators of estrogen over-exposure.
   

 Cancer Research. 2011, 71(19):6208-19. doi: 10.1158/0008-5472.CAN-11-0073.
 MicroRNA-708 induces apoptosis and suppresses tumorigenicity in renal cancer cells
 
 
 Saini S, Yamamura S, Majid S, Shahryari V, Hirata H, Tanaka Y, Dahiya R.
  Abstract
Cancer pathogenesis is restricted by stresses that compromise cell division and survival. In this study, we identify miR-708, a little studied member of a set of microRNAs that have been implicated in stress control, as an important tumor suppressor in renal cell carcinoma (RCC). miR-708 expression was attenuated widely in human RCC specimens. Restoration of miR-708 expression in RCC cell lines decreased cell growth, clonability, invasion, and migration and elicited a dramatic increase in apoptosis. Moreover, intratumoral delivery of miR-708 was sufficient to trigger in vivo regression of established tumors in murine xenograft models of human RCC. Investigation of the targets of miR-708 identified the inhibitor of apoptosis protein survivin as important. siRNA-mediated knockdown of survivin partially phenocopied miR-708 overexpression suggesting that the proapoptotic role of miR-708 may be mediated primarily through survivin regulation. Additionally, we identified the E-cadherin regulators ZEB2 and BMI1 as likely miR-708 targets. Taken together, our findings define a major tumor suppressive role for miR-708, which may offer an attractive new target for prognostic and therapeutic intervention in RCC.
   

 J Matern Fetal Neonatal Med. 2011, 24(8):1002-12. doi: 10.3109/14767058.2010.538454.
 Global maternal early pregnancy peripheral blood mRNA and miRNA expression profiles according to plasma 25-hydroxyvitamin D concentrations
 
 
 Enquobahrie DA, Williams MA, Qiu C, Siscovick DS, Sorensen TK.
  Abstract
We investigated associations of early pregnancy maternal vitamin D concentrations with differential gene expression and post-transcription regulation. Plasma 25-hydroxyvitamin D (25[OH]D) was measured among participants of a nested case-control study. Participants with low (<25.5 ng/ml) and high (?31.7 ng/ml) 25[OH]D were identified among controls. Peripheral blood messenger RNA (mRNA) (N?=?21) and microRNA (miRNA) (N?=?13) expression studies were conducted among participants with low and high 25[OH]D concentrations. Differential expression between low/high groups were evaluated using Student's t-test, fold change, and SAM comparisons. We further investigated functions and functional relationships of differentially expressed mRNAs and targets of differentially expressed miRNAs. Three hundred and five genes (299 upregulated and 6 downregulated) and 11 miRNAs (10 downregulated and 1 upregulated) were differentially expressed among participants with low 25[OH]D compared with those who had high 25[OH]D. Genes that participate in a wide range of cellular functions, including organ and system development (e.g. angiogenesis), inflammation and metabolic processes (e.g. carbohydrate/lipid metabolism), as well as miRNAs that target these genes were differentially expressed among women with low 25[OH]D compared with those with high 25[OH]D. Early pregnancy plasma 25[OH]D concentrations are associated with maternal peripheral blood gene expression and post-transcription regulation.
   

 Mutat Res. 2011, 707(1-2):42-52. doi: 10.1016/j.mrfmmm.2010.12.009.
 Antroquinonol inhibits NSCLC proliferation by altering PI3K/mTOR proteins and miRNA expression profiles
 
 
 Kumar VB, Yuan TC, Liou JW, Yang CJ, Sung PJ, Weng CF.
  Abstract
Antroquinonol a derivative of Antrodia camphorata has been reported to have antitumor effects against various cancer cells. However, the effect of antroquinonol on cell signalling and survival pathways in non-small cell lung cancer (NSCLC) cells has not been fully demarcated. Here we report that antroquinonol treatment significantly reduced the proliferation of three NSCLC cells. Treatment of A549 cells with antroquinonol increased cell shrinkage, apoptotic vacuoles, pore formation, TUNEL positive cells and increased Sub-G1 cell population with respect to time and dose dependent manner. Antroquinonol treatment not only increased the Sub-G1 accumulation but also reduced the protein levels of cdc2 without altering the expression of cyclin B1, cdc25C, pcdc2, and pcdc25C. Antroquinonol induced apoptosis was associated with disrupted mitochondrial membrane potential and activation of Caspase 3 and PARP cleavage in A549 cells. Moreover, antroquinonol treatment down regulated the expression of Bcl2 proteins, which was correlated with the decreased PI3K and mTOR protein levels without altering pro apoptotic and anti apoptotic proteins. Results from the microarray analysis demonstrated that antroquinonol altered the expression level of miRNAs compared with untreated control in A549 cells. The data collectively suggested the antiproliferative effect of antroquinonol on NSCLC A549 cells, which provides useful information for understanding the anticancer mechanism influenced by antroquinonol and is the first report to suggest that antroquinonol may be a promising chemotherapeutic agent for lung cancer.