Cytokeratin 8 Was Over-Expressed in Cells Harboring in Vitro-Transcribed Full Length Hepatitis C Virus 1b RNA, but Down-Expressed in HCV Patients’ Serum
Clinical Medicine Research
Volume 3, Issue 3, May 2014, Pages: 80-86
Received: May 21, 2014;
Accepted: Jun. 16, 2014;
Published: Jun. 30, 2014
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Meng Xun, Department of Immunology and Microbiology, Medical School of Xi’an Jiaotong University, Xi’an, Shannxi, China
Haifeng Wang, Department of Clinical Laboratories, Xi’an Central Hospital, Xi’an, Shannxi, China
Burong Li, Department of Clinical Laboratories, Second Affiliated Hospital of Medical School of Xi’an Jiaotong University, Xi’an, Shannxi, China
Hongyan He, Department of Medical Technology, Xi’an Medical College, Xi’an, Shannxi, China
Qian He, Department of Clinical Laboratories, Second Affiliated Hospital of Medical School of Xi’an Jiaotong University, Xi’an, Shannxi, China
Yonglie Chu, Department of Immunology and Microbiology, Medical School of Xi’an Jiaotong University, Xi’an, Shannxi, China
Objectives: Previous proteome analysis conducted by our group has demonstrated that cytokeratin 8 was overexpressed in HuH-7 cells harboring in vitro transcribed HCV 1b full length RNA (HuH-7-HCV). Present study was aim to verify the results of proteomics, and obtain the clinical data of CK8 expresson levels in HCV patients. Methods: The expression level of CK 8 in HuH-7-HCV cells was examined by Real time PCR and Western blotting. The concentration of CK8 in HCV patients’ serum was measured by enzyme-linked immunosorbent assay (ELISA). Results: The results showed expression level of CK8 transcript in HuH-7-HCV cells was 2.3 fold higher than that in HuH-7 mock cells (P<0.01). The protein expression of CK8 in HuH-7-HCV cells was approximately 3 fold higher than that in HuH-7 mock cells (P<0.01). However, results of ELISA demonstrated the serum CK8 concentration was significantly reduced in chronic HCV patients compared to normal healthy controls (P<0.01). And there was a negative linear correlation between serum CK8 concentration and HCV RNA titer (r=-0.380, P<0.01). Conclusion: Our present study supports the hypothesis that in response to HCV infection, expression of CK8 was increased, which may contribute to the essential cytoprotection provided by CK8 in the liver. Altered CK8 expression pattern could be an important event in the pathogenesis of HCV infection. CK8 have potential use as surrogate markers of liver injury.
Cytokeratin 8 Was Over-Expressed in Cells Harboring in Vitro-Transcribed Full Length Hepatitis C Virus 1b RNA, but Down-Expressed in HCV Patients’ Serum, Clinical Medicine Research.
Vol. 3, No. 3,
2014, pp. 80-86.
Ezzikouri, S., P. Pineau and S. Benjelloun, Hepatitis C virus infection in the Maghreb region. J Med Virol, 2013. 85(9): p. 1542-9.
Lagging, M., et al., Retreatment with peg-interferon and ribavirin in patients with chronic hepatitis C virus genotype 2 or 3 infection with prior relapse. Scand J Gastroenterol, 2013. 48(7): p. 839-47.
Xun, M., et al., Proteomic analysis of HuH-7 cells harboring in vitro-transcribed full-length hepatitis C virus 1b RNA. Acta Pharmacol Sin, 2008. 29(6): p. 720-7.
Tang, H.L., et al., inventors; Yangling, Daiying Biolog Engine (CN), as-signee. The intact hepatitis C virus and the method for culturing it in in vitro cell culture. European Patent 1 424 390. 2004 Feb 06.
Livak, K.J. and T.D. Schmittgen, Analysis of relative gene expression data using real-time quan-titative PCR and the 2(-Delta Delta C(T)) Method. Methods, 2001. 25(4): p. 402-8.
Herrmann, H., et al., In-termediate filaments: primary determinants of cell architecture and plasticity. J Clin Invest, 2009. 119(7): p. 1772-83.
Strnad, P., et al., Non-coding keratin variants associate with liver fibrosis progression in patients with hemochromatosis. PLoS One, 2012. 7(3): p. e32669.
Omary, M.B., "IF-pathies": a broad spectrum of intermediate filament-associated diseases. J Clin Invest, 2009. 119(7): p. 1756-62.
Iyer, S.V., et al., Under-standing the role of keratins 8 and 18 in neoplastic potential of breast cancer derived cell lines. PLoS One, 2013. 8(1): p. e53532.
Usachov, V., et al., Keratin 8 variants are infrequent in patients with alcohol-related liver cirrhosis and do not associate with development of hepatocellular carcinoma. BMC Gastroenterol, 2012. 12: p. 147.
Fortier, A.M., et al., Novel insights into changes in biochemical properties of keratins 8 and 18 in gri-seofulvin-induced toxic liver injury. Exp Mol Pathol, 2010. 89(2): p. 117-25.
Omary, M.B., et al., Toward un-raveling the complexity of simple epithelial keratins in human disease. J Clin Invest, 2009. 119(7): p. 1794-805.
Ku, N.O., et al., Keratins let liver live: Mutations predispose to liver disease and crosslinking ge-nerates Mallory-Denk bodies. Hepatology, 2007. 46(5): p. 1639-49.
Zatloukal, K., et al., From Mallory to Mallory-Denk bodies: what, how and why? Exp Cell Res, 2007. 313(10): p. 2033-49.
Kakehashi, A., et al., Cytokeratin 8/18 overexpression and complex formation as an indicator of GST-P positive foci transformation into hepatocellular carcinomas. Toxicol Appl Pharmacol, 2009. 238(1): p. 71-9.
Kakehashi, A., et al., Cytokeratin 8/18 as a new marker of mouse liver preneoplastic lesions. Toxicol Appl Pharmacol, 2010. 242(1): p. 47-55.
Zhong, B., et al., Keratin variants are overrepresented in primary biliary cirrhosis and associate with disease severity. Hepatology, 2009. 50(2): p. 546-54.
Strnad, P., et al., Keratins: markers and modulators of liver disease. Curr Opin Gastroenterol, 2012. 28(3): p. 209-16.
Strnad, P., et al., Keratin variants associate with progression of fibrosis during chronic hepatitis C infection. Hepatology, 2006. 43(6): p. 1354-63.
Lee, J., et al., Predisposition to apoptosis in keratin 8-null liver is related to inactivation of NF-kappaB and SAPKs but not decreased c-Flip. Biol Open, 2013. 2(7): p. 695-702.
Wang, Y., et al., Cytokeratin 8 silencing in human naso-pharyngeal carcinoma cells leads to cisplatin sensitization. Cancer Lett, 2008. 265(2): p. 188-96.
Caulin, C., et al., Keratin-dependent, epithelial resistance to tumor necrosis factor-induced apoptosis. J Cell Biol, 2000. 149(1): p. 17-22.
Gilbert, S., et al., Simple epithelium keratins 8 and 18 provide resistance to Fas-mediated apoptosis. The protection occurs through a receptor-targeting modulation. J Cell Biol, 2001. 154(4): p. 763-73.
Fuentes-Gonzalez, A.M., et al., The modulation of apoptosis by oncogenic viruses. Virol J, 2013. 10: p. 182.
Bantel, H. and K. Schulze-Osthoff, Apoptosis in hepatitis C virus infection. Cell Death Differ, 2003. 10 Suppl 1: p. S48-58.
Valva, P., et al., Apoptosis markers related to pathogenesis of pediatric chronic hepatitis C virus infection: M30 mirrors the severity of steatosis. J Med Virol, 2010. 82(6): p. 949-57.