Hsa-miR-331-3p inhibits VHL expression by directly targeting its mRNA 3'-UTR in HCC cell lines

• Authors: Yiyi Cao , Jinnan Zhang , Dongmei Xiong , Dan Wang , Ting Wu , Ailong Huang , Hua Tang
• Languages of publication: EN

Abstracts

EN

Dysregulation of miRNA is widely involved in human cancers, including hepatocellular carcinoma (HCC). Array data for miRNAs indicated that miR-331-3p might be one of the disorderly expressed miRNAs in HCC cell lines, but the function of miR-331-3p in HCC remains unclear. In this study, quantitative real time polymerase chain reaction (qRT-PCR) results indicated that miR-331-3p was up-regulated in HepG2.2.15 cells, Ad-HBV-HepG2 cells and pCH9/3091transfected SMMC7721 cells compared with their control group, respectively. miRNA target prediction software was used, and VHL was found to be one of the target genes of miR-331-3p. qRT-PCR and western blot analysis indicated VHL expression was decreased when miR-331-3p was over-expressed and increased when miR-331-3p was inhibited in SMMC7721 cells. The luciferase reporter activity was inhibited in SMMC7721 cells when co-transfected with miR-331-3p expression vector and VHL 3'-UTR wild type vector and increased in HepG2.2.15 transfected with miR-331-3p inhibitor compared to its control group respectively. When co-transfected with miR-331-3p expression vector and VHL 3'-UTR mutated type vector in SMMC7721 cells the luciferase reporter activity was recovered. All of these results show that HBV up-regulated miR-331-3p expression in HCC cell lines and miR-331-3p could inhibit VHL expression by directly targeting its 3'-UTR. This provided useful information in exploring the mechanism of HCC induced by HBV infection.

Keywords

EN

HCC; HBV; miRNA; Hsa-miR-331-3p and VHL

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2015
• Volume: 62
• Issue: 1
• Pages: 77-82

Dates

published

2015

received

2014-04-23

revised

2014-12-05

accepted

2015-01-09

online

2015-02-26

Contributors

author

Yiyi Cao

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

author

Jinnan Zhang

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

author

Dongmei Xiong

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

author

Dan Wang

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

author

Ting Wu

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

author

Ailong Huang

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

author

Hua Tang

• Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, Chongqing Medical University, Chongqing 400016, China

References

• Ambros V (2004) The functions of animal microRNAs. Nature 431: 350-355.
• Bartel D (2004) MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell 116: 281-297.
• Calin GA, Croce CM (2006) MicroRNA signatures in human cancers. Nat Rev Cancer 6: 857-866.
• Cockman ME, Masson N, Mole DR, Jaakkola P, Chang GW, Clifford SC, Maher ER, Pugh CW, Ratcliffe PJ, Maxwell PH. (2000) Hypoxia inducible factor-alpha binding and ubiquitylation by the von Hippel-Lindau tumor suppressor protein. J Biol Chem 275: 25733-25741.
• Croce CM (2009) Causes and consequences of micro-RNA dysregulation in cancer. Nat Rev Genet 10: 704-714.
• Epis MR, Barker A, Giles KM, Beveridge DJ, Leedman PJ (2011) The RNA-binding protein HuR opposes the repression of ERBB-2 gene expression by microRNAmiR-331-3p in prostate cancer cells. J Biol Chem 286: 41442-41454.
• Epis MR, Giles KM, Candy PA, Webster RJ, Leedman PJ (2014) MiR-331-3p regulates expression of neuropilin-2 in glioblastoma. J Neurooncol 116: 67-75.
• Epis MR, Giles KM, Kalinowski FC, Barker A, Cohen RJ, Leedman PJ (2012) Regulation of expression of deoxyhypusine hydroxylase (DOHH), the enzyme that catalyzes the activation of eIF5A, by miR-331-3p and miR-642-5p in prostate cancer cells. J Biol Chem 287: 35251-35259.
• Gaál Z, Oláh E (2012) MicroRNAs and their role in malignant hematologic diseases. Orv Hetil 153: 2051-2059.
• Gao P, Wong CC, Tung EK, Lee JM, Wong CM, Ng IO (2011) Deregulation of microRNA expression occurs early and accumulates in early stages of HBV-associated multistep hepatocarcinogenesis. J Hepatol 54: 1177-1184.
• Griffiths-Jones S, Saini HK, van Dongen S, Enright AJ (2008) MiRBase: tools for microRNA genomics. Nucleic Acids Res 36: D154-D158.
• Guo X, Guo L, Ji J, Zhang J, Zhang J, Chen X, Cai Q, Li J, Gu Q, Liu B, Zhu Z, Yu Y (2010) MiRNA-331-3p directly targets E2F1 and induces growth arrest in human gastric cancer. Biochem Biophys Res Commun 398: 1-6.
• Kim WY, Kaelin WG (2004) Role of VHL gene mutation in human cancer. J Clin Oncol 22: 4991-5004.
• Latif F, Tory K, Gnarra J, Yao M, Duh FM, Orcutt ML, Stackhouse T, Kuzmin L, Modi W, Geil L (1993) Identification of the von Hippel-Lindau disease tumor suppressor gene. Science 260: 1317-1320.
• Lin S, Pan L, Guo S, Wu J, Jin L, Wang JC, Wang S (2013) Prognostic role of microRNA-181a/b in hematological malignancies: a meta-analysis. PLoS One 8: e59532.
• Lin Z, Flemington EK (2011) MiRNAs in the pathogenesis of oncogenic human viruses. Cancer Lett 305: 186-199.
• Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−ΔΔC(T)) method. Methods 25: 402-408.
• Luo J, Deng ZL, Luo X, Tang N, Song WX, Chen J, Sharff KA, Luu HH, Haydon RC, Kinzler KW, Vogelstein B, He TC (2007) A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. Nat Protoc 2: 1236-1247.
• Maher ER, Kaelin WG Jr (1997) Von Hippel-Lindau disease. Medicine (Baltimore) 76: 381-391.
• Wang CM, Wang Y, Fan CG, Xu FF, Sun WS, Liu YG, Jia JH (2011) MiR-29c targets TNFAIP3, inhibits cells proliferation and induces apoptosis in hepatitis B virus-related hepatocellular carcinoma. Biochem Biophys Res Commun 411: 586-592.
• Xu X, Fan Z, Kang L, Han J, Jiang C, Zheng X, Zhu Z, Jiao H, Lin J, Jiang K, Ding L, Zhang H, Cheng L, Fu H, Song Y, Jiang Y, Liu J, Wang R, Du N, Ye Q (2013) Hepatitis B virus X protein represses miRNA-148a to enhance tumorigenesis. J Clin Invest 123: 630-645.
• Yip WK, Cheng AS, Zhu R, Lung RW, Tsang DP, Lau SS, Chen Y, Sung JG, Lai PB, Ng EK, Yu J, Wong N, To KF, Wong VW, Sung JJ, Chan HL (2011) Carboxyl-terminal truncated HBx regulates a distinct microRNA transcription program in hepatocellsular carcinoma development. PLoS One 6: e22888.
• Zhang B, Pan X, Cobb GP, Anderson TA (2007) microRNAs as oncogenes and tumor suppressors. Dev Biol 302: 1-12.
• Zhang ZZ, Liu X, Wang DQ, Teng MK, Niu LW, Huang AL (2011) Hepatitis B virus and hepatocelluar carcinoma at the mRNA level. World J Gastroenterol 17: 3353-3358.
• Zou CC, Li YG, Cao YY, Zhang JN, Jiang JR, Sheng YR, Wang S, Huang AL, Tang H (2014) Up-regulated MicroRNA-181a induces carcinogenesis in Hepatitis B virus-related hepatocellular carcinoma by targeting E2F5. BMC Cancer 14: 97.