Hepatitis C virus NS3 protein modulates the biological behaviors of malignant hepatocytes by altering the expression of host cell microRNA

Zhang,Jun; Ishigaki,Yasuhito; Takegami,Tsutomu

doi:10.3892/mmr.2015.4041

October-2015 Volume 12 Issue 4

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October-2015 Volume 12 Issue 4

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Article

Hepatitis C virus NS3 protein modulates the biological behaviors of malignant hepatocytes by altering the expression of host cell microRNA

Authors:
- Jun Zhang
- Yasuhito Ishigaki
- Tsutomu Takegami
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Affiliations: Department of Comprehensive Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, P.R. China, Department of Life Science, Medical Research Institute, Kanazawa Medical University, Uchinada, Ishikawa 920‑0293, Japan
Pages: 5109-5115
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Published online on: July 3, 2015

https://doi.org/10.3892/mmr.2015.4041
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Abstract

Chronic hepatitis C virus (HCV) infection is a major cause of hepatocellular carcinoma (HCC) worldwide. The HCV non‑structural protein 3 (NS3) protein is considered to affect normal cellular functions and to be involved in HCV carcinogenesis. The expression of microRNA (miRNA) is altered in human HCC, thus implicating its role in hepatocarcinogenesis. To investigate the mechanisms by which the HCV NS3 protein affects the expression of miRNA in malignant hepatocytes, if any, the present study constructed expression vectors encoding the HCV NS3 and NS3/4A proteins, which were stably transfected into HepG2 cells. The biological behaviors of the HepG2 transfectants and their differential expression levels of miRNA expression were investigated. Compared with the HepG2‑vector cells, the HepG2‑NS3 cells grew at a slower rate, were arrested in the G0/G1 cell cycle phase, formed more colonies and developed larger tumors at a faster rate. Co‑expression of HCV NS4A resulted in the inhibition of HCV NS3‑stimulated tumorigenicity. A total of 35 miRNAs were dysregulated, 26 of which were downregulated and nine of which were upregulated, in the HepG2‑NS3 cells, and 75 miRNAs were altered in HepG2‑NS3/4A cells, of which 20 were downregulated and 55 were upregulated). In addition, significant decreases in the mRNA levels of p53 and p21 were observed, which confirmed differential expression of miRNA. These results suggested that differential miRNA profiling in malignant hepatocytes may account for the variable pathophysiological manifestations associated with the HCV NS3 protein. These differentially expressed miRNAs may offer potential as candidates for the development of miRNA‑based therapeutics.

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1	Koike K, Tsutsumi T, Fujie H, Shintani Y and Kyoji M: Molecular mechanism of viral hepatocarcinogenesis. Oncology. 62(Suppl 1): 29–37. 2002. View Article : Google Scholar : PubMed/NCBI
2	Takamizawa A, Mori C, Fuke I, et al: Structure and organization of the hepatitis C virus genome isolated from human carriers. J Virol. 65:1105–1113. 1991.PubMed/NCBI
3	Neddermann P, Tomei L, Steinkühle C, Gallinari P, Tramontano A and De Francesco R: The nonstructural proteins of the hepatitis C virus: structure and functions. Biol Chem. 378:469–476. 1997.PubMed/NCBI
4	Penin F, Dubuisson J, Rey FA, Moradpour D and Pawlotsky JM: Structural biology of hepatitis C virus. Hepatology. 39:5–19. 2004. View Article : Google Scholar : PubMed/NCBI
5	Grakoui A, Levis R, Raju R, Huang HV and Rice CM: A cis-acting mutation in the sindbis virus junction region which affects subgenomic RNA synthesis. J Virol. 63:5216–5227. 1989.PubMed/NCBI
6	Hong Z, Ferrari E, Wright-Minogue J, et al: Enzymatic characterization of hepatitis C virus NS3/4A complexes expressed in mammalian cells by using the herpes simplex virus amplicon system. J Virol. 70:4261–4268. 1997.
7	Suzich JA, Tamura JK, Palmer-Hill F, et al: Hepatitis C virus NS3 protein polynucleotide-stimulated nucleotide triphosphatase and comparison with the related pestivirus and flavivirus enzymes. J Virol. 67:6152–6158. 1993.PubMed/NCBI
8	Kim DW, Gwack Y, Han JH and Choe J: C-terminal domain of the hepatitis C virus NS3 protein contains an RNA helicase activity. Biochem Biophys Res Commun. 215:160–166. 1995. View Article : Google Scholar : PubMed/NCBI
9	Sakamuro D, Furukawa T and Takegami T: Hepatitis C virus nonstructural protein NS3 transforms NIH 3T3 cells. J Virol. 69:3893–3896. 1995.PubMed/NCBI
10	He QQ, Cheng RX, Sun Y, Feng DY, Chen ZC and Zheng H: Hepatocyte transformation and tumor development induced by hepatitis C virus NS3 c-terminal deleted protein. World J Gastroenterol. 9:474–478. 2003. View Article : Google Scholar : PubMed/NCBI
11	Carthew RW and Sontheimer EJ: Origins and mechanisms of miRNAs and siRNAs. Cell. 136:642–655. 2009. View Article : Google Scholar : PubMed/NCBI
12	Jopling CL, Yi M, Lancaster AM, Lemon SM and Sarnow P: Modulation of hepatitis C virus RNA abundance by a liver-specific MicroRNA. Science. 309:1577–1581. 2005. View Article : Google Scholar : PubMed/NCBI
13	Hou W, Tian Q, Zheng J and Bonkovsky HL: MicroRNA-196 represses bach1 protein and hepatitis C virus gene expression in human hepatoma cells expressing hepatitis C viral proteins. Hepatology. 51:1494–1504. 2010. View Article : Google Scholar : PubMed/NCBI
14	Murakami Y, Yasuda T, Saigo K, et al: Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues. Oncogene. 25:2537–2545. 2006. View Article : Google Scholar
15	Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST and Patel T: MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology. 133:647–658. 2007. View Article : Google Scholar : PubMed/NCBI
16	Gramantieri L, Ferracin M, Fornari F, et al: Cyclin G1 is a target of miR122a, a microRNA frequently down-regulated in human cholangiocarcinoma. Cancer Res. 67:6092–6099. 2007. View Article : Google Scholar : PubMed/NCBI
17	Hammond JB and Kruger NJ: The Bradford method for protein quantitation. Methods Mol Biol. 3:25–32. 1988.PubMed/NCBI
18	Livak JK and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar
19	EI-Serag HB, Marrero JA, Rudolph L and Reddy KR: Diagnosis and treatment of hepatocellular carcinoma. Gastroenterology. 134:1752–1763. 2008. View Article : Google Scholar
20	Kasprzak A and Adamek A: Role of hepatitis C virus proteins (C, NS3, NS5A) in hepatic oncogenesis. Hepatol Res. 38:1–26. 2008. View Article : Google Scholar
21	Moriya K, Fujie H, Shintani Y, et al: The core protein of hepatitis C virus induces hepatocellular carcinoma in transgenic mice. Nat Med. 4:1065–1067. 1998. View Article : Google Scholar : PubMed/NCBI
22	Lerat H, Honda M, Beard MR, et al: Steatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus. Gastroenterology. 122:352–365. 2002. View Article : Google Scholar : PubMed/NCBI
23	Kwun HJ, Jung EY, Ahn JY, Lee MN and Jang KL: p53-dependent transcriptional repression of p21 (waf1) by hepatitis C virus NS3. J Gen Virol. 82:2235–2241. 2001. View Article : Google Scholar : PubMed/NCBI
24	Siavoshian S, Abraham JD, Kieny MP and Schuster C: HCV core, NS3, NS5A and NS5B proteins modulate cell proliferation independently from p53 expression in hepatocarcinoma cell lines. Arch Virol. 149:323–336. 2004. View Article : Google Scholar : PubMed/NCBI
25	Hassan M, Ghozlan H and Abdel-Kader O: Activation of c-Jun NH2-terminal kinase (JNK) signaling pathway is essential for the stimulation of hepatitis C virus (HCV) non-structural protein 3 (NS3)-mediated cell growth. Virology. 333:324–336. 2005. View Article : Google Scholar : PubMed/NCBI
26	Randall G, Panis M, Cooper JD, et al: Cellular cofactors affecting hepatitis C virus infection and replication. Proc Natl Acad Sci USA. 104:12884–12889. 2007. View Article : Google Scholar : PubMed/NCBI
27	Landgraf P, Rusu M, Sheridan R, et al: A mammalian microRNA expression atlas based on small RNA library sequencing. Cell. 129:1401–1414. 2007. View Article : Google Scholar : PubMed/NCBI
28	Wang Y, Kato N, Jazag A, et al: Hepatitis C virus core protein is a potent inhibitor of RNA silencing-based antiviral response. Gastroenterology. 130:883–892. 2006. View Article : Google Scholar : PubMed/NCBI
29	Lipardi C, Wei Q and Paterson BM: RNAi as random degradative PCR: siRNA primers convert mRNA into dsRNAs that are degraded to generate new sirNAs. Cell. 107:297–307. 2001. View Article : Google Scholar : PubMed/NCBI
30	Kutay H, Bai S, Datta J, et al: Downregulation of miR-122 in the rodent and human hepatocellular carcinomas. J Cell Biochem. 99:671–678. 2006. View Article : Google Scholar : PubMed/NCBI
31	Ishido S, Fujita T and Hotta H: Complex formation of NS5B with NS3 and NS4A proteins of hepatitis C virus. Biochem Biophys Res Commun. 244:35–40. 1998. View Article : Google Scholar : PubMed/NCBI
32	Yoon AR, Gao R, Kaul Z, et al: MicroRNA-296 is enriched in cancer cells and downregulates p21WAF1 mRNA expression via interaction with its 3′ untranslated region. Nucleic Acids Res. 39:8078–8091. 2011. View Article : Google Scholar : PubMed/NCBI