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2013, Number 5

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Ann Hepatol 2013; 12 (5)

Use of proteomic analysis tools to identify HCV-proteins down-regulated by acetylsalicylic acid

Sánchez-García A, Ríos-Ibarra CP, Rincón-Sánchez AR, Ortiz-López R, Garza-Juárez A, Morlett-Chávez J, Martínez-Rodríguez H, Rivas-Estilla AM

Language: English
References: 28
Page: 725-732
PDF size: 319.96 Kb.


ABSTRACT

Background and aim. Acetylsalicylic acid (ASA) has been shown to downregulate HCV expression; however, the involved mechanisms are unknown. We used proteomic analysis to compare protein expression profiles between human hepatocarcinoma cells (Huh7) and Huh7-HCV cells harboring expression of non-structural HCV proteins, to elucidate the mechanism(s) involved in ASA-mediated downregulation of HCV replication. Material and methods. Both cell lines were treated or untreated with 4 mM ASA and harvested at 0, 24, 48 and 72 h to isolate total proteins, which were resolved by two-dimensional gel electrophoresis (2DE) to separate them by isoelectric point (pI), followed by fractionation by molecular weight (MW). Gels were scanned and analyzed with PD-Quest software V8.0.1, and proteins were elucidated by the specific pI and MW using TAGIDENT software. Statistics analysis included the t-test. Results and Discussion. Different protein patterns among hepatocytes expressing HCV-proteins in ASA treated and untreated cells were found. Among proteins differentially expressed in Huh7-HCV cells, we found proteins related to cell proliferation (MTMR6, FAM22, HDGF and HCF-1) after 24 h of ASA treatment; and upregulation of angiostatin, PI4KA and STAT-1 after 48 h of treatment. Finally, at 72 h of ASA exposure, we identified overexpression of adenylsuccinate synthase, 2’-3’-di-deoxyadenosine, ubiquitin-protein-ligase E6A, adenylosuccinate-lyase and nibrin (NBN). Conclusion. We found that ASA induces different protein patterns in Huh7-HCV cells promoting activation of proteins involved in cell progression, repair of double strand breaks, proliferation, inhibition of apoptosis and growth stimulation at the same time that it decreased HCV expression.


REFERENCES

  1. Alter M. Epidemiology of hepatitis C virus infection. World J Gastroenterol 2007; 13: 2436-41.

  2. Drazan K. Molecular biology of hepatitis C infection. Liver Transplantation 2000; 6: 396-406.

  3. Sabahi A. Hepatitis C virus entry: the early stops in the viral replication cycle. Virology J 2009; 6: 117.

  4. Sy T, Jamal M. Epidemiology of Hepatitis C virus (HCV) infection. Int J Med Sci 2006; 3: 41-6.

  5. Brass V, Moradpour D, Blum H. Molecular virology of hepatitis C virus (HCV): 2006 Update. Int J Medical Sciences 2006; 3: 29-34.

  6. Chevaliez S, Pawlotsky J. Hepatitis C virus: virology, diagnosis and management of antiviral therapy. World J Gastroenterol 2007; 13: 2461-6.

  7. Blight K, Kolykhalov A, Rice C. Efficient initiation of HCV RNA replication in cell culture. Science 2000; 290: 1972-4.

  8. Lindenbach BD, Evans MJ, Syder AJ, Wolk B, Tellinghuisen TL, Liu CC, Maruyama T, et al. Complete replication of hepatitis C virus in cell culture. Science 2005; 309: 623-6.

  9. Lohmann V, Korner F, Koch J, Herian U, Theilmann L, Bartenschlager R. Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 1999; 285: 110-3.

  10. Wakita T, Pietschmann T, Kato T, Date T, Miyamoto M, Zhao Z, Murthy K, et al. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med 2005; 11: 791-6.

  11. Trujillo K, Rincón AR, Martínez H, Bosques F, Ramos J, Barrera HA, Rivas AM, et al. Acetylsalicylic acid inhibits hepatitis C virus RNA and protein expression through cyclooxygenase-2 signaling pathways. Hepatology 2008; 47: 1462-72.

  12. Jacobs JM, Diamond DL, Chan EY, Gritsenko MA, Qian W, Stastna M, Baas T, et al. Proteome analysis of liver cells expressing a full length hepatitis C virus (HCV) replicon and biopsy specimens of posttransplantation liver from HCV-infected patients. J Virol 2005; 79: 7558-9.

  13. Castagnino J. Proteómica y sus aplicaciones en medicina humana. Acta Bioquim Clin Latinoam 2008; 42: 179-82.

  14. Hortin G, Jortani S, Ritchie J, Valdes R, Chan D. Proteomics: A new diagnostic frontier. Clinical Chemistry 2006; 52: 1218-22.

  15. Molloy M, Witzmann F. Proteomics: Technologies and applications. Briefings in functional genomics and proteomics 2001; 1: 23-9.

  16. Motonari T, Yasuhiro K, Yuuichiro Y, Norio I, Toshifusa T, Isao S, Kiwamu O, et al. Proteomic profiling of heat shock protein 70 family members as biomarkers for hepatitis C virus-related hepatocellular carcinoma. Proteomics 2003; 3: 2487-93.

  17. Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A. Protein identification and analysis tools on the ExPASy Server. In: Walker JM (ed). NJ: Humana Press Inc., 2005, p 571-607.

  18. Liao CL, Lin YL, Wu BC, Tsao CH, Wang MC, Liu CI, Huang YL, et al. Salicylates inhibit flavivirus replication independently of blocking nuclear factor kappa B activation. J Virol 2001; 75: 7828-39.

  19. Speir E, Yu ZX, Ferrans VJ, Huang ES, Epstein SE. Aspirin attenuates cytomegalovirus infectivity and gene expression mediated by cyclooxygenase-2 in coronary artery smooth muscle cells. Circ Res 1998; 83: 210-6.

  20. Primache V, Binda S, De Benedittis G, Barbi M. In vitro activity of acetylsalicylic acid on replication of varicella-zoster virus. New Microbiol 1998; 21: 397-401.

  21. Xun M, Zhao SH, Cao CX, Song J, Shao MM, Chu YL. Proteomic analysis of Huh-7 cells harboring in Vitro-transcribed full-length hepatitis C virus 1b RNA. Act Pharmacol Sin 2008; 29: 720-7.

  22. Diamond DL, Jacobs JM, Paeper B, Proll SC, Gritsenko MA, Carithers RL, Larson AM, et al. Proteomic profiling of human liver biopsies: hepatitis C virus-induced fibrosis and mitochondrial dysfunction. Hepatology 2007; 46: 649-57.

  23. Joyce MA, Walters KA, Lamb SE, Yeh MM, Zhu LF, Kneteman N, Doyle JS, et al. HCV induces oxidative and ER stress, and sensitizes infected cells to apoptosis in SCID/ Alb-uPA mice. PLoS Pathog 2009; 5: e1000291.

  24. Fang C, Yi Z, Liu F, Lan S, Wang J, Lu H, Yang P, et al. Proteome analysis of human liver carcinoma Huh7 cells harboring hepatitis C virus subgenomic replicon. Proteomics 2006; 6: 519-27.

  25. Hartman SE, Bertone P, Nath AK, Royce TE, Gerstein M, Weissman S, Snyder M. Global changes in STAT target selection and transcription regulation upon interferon treatments. Genes Dev 2005; 19: 2953-68.

  26. Hinze K, Wald K, Zeng H, Jeffery E, Finley J. Thioredoxin reductase in human hepatoma cells is transcriptionally regulated by sulforaphane and other electrophiles via an antioxidant response element. J Nutr 2003; 133: 2721-7.

  27. Ganther HE. Selenium metabolism, selenoproteins and mechanisms of cancer prevention: complexities with thioredoxin reductase. Carcinogenesis 1999; 20: 1657-66.

  28. Shirakura M, Murakami K, Ichimura T, Suzuki R, Shimoji T, Fukuda K, Abe K, et al. E6AP ubiquitin ligase mediates ubiquitylation and degradation of hepatitis C virus core protein. J Virol 2007; 81: 1174-85.




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