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Published in: Virology Journal 1/2011

Open Access 01-12-2011 | Research

Hepatitis B virus induces G1 phase arrest by regulating cell cycle genes in HepG2.2.15 cells

Authors: Tianzhen Wang, Ran Zhao, Yiqi Wu, Dan Kong, Lei Zhang, Di Wu, Chao Li, Chong Zhang, Zuxi Yu, Xiaoming Jin

Published in: Virology Journal | Issue 1/2011

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Abstract

Background

To investigate the effect of HBV on the proliferative ability of host cells and explore the potential mechanism.

Methods

MTT, colony formation assay and tumourigenicity in nude mice were performed to investigate the effect of HBV on the proliferative capability of host cells. In order to explore the potential mechanism, cell cycle and apoptosis were analysed. The cell cycle genes controlling the G1/S phase transition were detected by immunohistochemistry, westernblot and RT-PCR.

Results

HepG2.2.15 cells showed decreased proliferation ability compared to HepG2 cells. G1 phase arrest was the main cause but was not associated with apoptosis. p53, p21 and total retinoblastoma (Rb) were determined to be up-regulated, whereas cyclinE was down-regulated at both the protein and mRNA levels in HepG2.2.15 cells. The phosphorylated Rb in HepG2.2.15 cells was decreased.

Conclusions

Our results suggested that HBV inhibited the capability of proliferation of HepG2.2.15 cells by regulating cell cycle genes expression and inducing G1 arrest.
Appendix
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Literature
1.
go back to reference Beasley RP, Hwang LY, Lin CC, Chien CS: Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet 1981, 2: 1129-1133.CrossRefPubMed Beasley RP, Hwang LY, Lin CC, Chien CS: Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet 1981, 2: 1129-1133.CrossRefPubMed
2.
go back to reference Dalgleish AG, Woods RL, Levi JA, Raghavan D, McCaughan GW, Tattersall MH: The role of hepatitis B virus in the etiology of hepatocellular carcinoma in Australia. Aust N Z J Med 1983, 13: 605-607.CrossRefPubMed Dalgleish AG, Woods RL, Levi JA, Raghavan D, McCaughan GW, Tattersall MH: The role of hepatitis B virus in the etiology of hepatocellular carcinoma in Australia. Aust N Z J Med 1983, 13: 605-607.CrossRefPubMed
3.
go back to reference Perz JF, Armstrong GL, Farrington LA, Hutin YJ, Bell BP: The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide. J Hepatol 2006, 45: 529-538. 10.1016/j.jhep.2006.05.013CrossRefPubMed Perz JF, Armstrong GL, Farrington LA, Hutin YJ, Bell BP: The contributions of hepatitis B virus and hepatitis C virus infections to cirrhosis and primary liver cancer worldwide. J Hepatol 2006, 45: 529-538. 10.1016/j.jhep.2006.05.013CrossRefPubMed
4.
go back to reference Wang Y, Wu MC, Sham JS, Tai LS, Fang Y, Wu WQ, Xie D, Guan XY: Different expression of hepatitis B surface antigen between hepatocellular carcinoma and its surrounding liver tissue, studied using a tissue microarray. J Pathol 2002, 197: 610-616. 10.1002/path.1150CrossRefPubMed Wang Y, Wu MC, Sham JS, Tai LS, Fang Y, Wu WQ, Xie D, Guan XY: Different expression of hepatitis B surface antigen between hepatocellular carcinoma and its surrounding liver tissue, studied using a tissue microarray. J Pathol 2002, 197: 610-616. 10.1002/path.1150CrossRefPubMed
5.
go back to reference Chen BF, Liu CJ, Jow GM, Chen PJ, Kao JH, Chen DS: High prevalence and mapping of pre-S deletion in hepatitis B virus carriers with progressive liver diseases. Gastroenterology 2006, 130: 1153-1168. 10.1053/j.gastro.2006.01.011CrossRefPubMed Chen BF, Liu CJ, Jow GM, Chen PJ, Kao JH, Chen DS: High prevalence and mapping of pre-S deletion in hepatitis B virus carriers with progressive liver diseases. Gastroenterology 2006, 130: 1153-1168. 10.1053/j.gastro.2006.01.011CrossRefPubMed
6.
go back to reference Liu CJ, Chen BF, Chen PJ, Lai MY, Huang WL, Kao JH, Chen DS: Role of hepatitis B virus precore/core promoter mutations and serum viral load on noncirrhotic hepatocellular carcinoma: a case-control study. J Infect Dis 2006, 194: 594-599. 10.1086/505883CrossRefPubMed Liu CJ, Chen BF, Chen PJ, Lai MY, Huang WL, Kao JH, Chen DS: Role of hepatitis B virus precore/core promoter mutations and serum viral load on noncirrhotic hepatocellular carcinoma: a case-control study. J Infect Dis 2006, 194: 594-599. 10.1086/505883CrossRefPubMed
7.
go back to reference Luan F, Liu H, Gao L, Liu J, Sun Z, Ju Y, Hou N, Guo C, Liang X, Zhang L, et al.: Hepatitis B virus protein preS2 potentially promotes HCC development via its transcriptional activation of hTERT. Gut 2009, 58: 1528-1537. 10.1136/gut.2008.174029CrossRefPubMed Luan F, Liu H, Gao L, Liu J, Sun Z, Ju Y, Hou N, Guo C, Liang X, Zhang L, et al.: Hepatitis B virus protein preS2 potentially promotes HCC development via its transcriptional activation of hTERT. Gut 2009, 58: 1528-1537. 10.1136/gut.2008.174029CrossRefPubMed
8.
go back to reference Slagle BL, Zhou YZ, Butel JS: Hepatitis B virus integration event in human chromosome 17p near the p53 gene identifies the region of the chromosome commonly deleted in virus-positive hepatocellular carcinomas. Cancer Res 1991, 51: 49-54.PubMed Slagle BL, Zhou YZ, Butel JS: Hepatitis B virus integration event in human chromosome 17p near the p53 gene identifies the region of the chromosome commonly deleted in virus-positive hepatocellular carcinomas. Cancer Res 1991, 51: 49-54.PubMed
9.
go back to reference Sells MA, Chen ML, Acs G: Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci USA 1987, 84: 1005-1009. 10.1073/pnas.84.4.1005PubMedCentralCrossRefPubMed Sells MA, Chen ML, Acs G: Production of hepatitis B virus particles in Hep G2 cells transfected with cloned hepatitis B virus DNA. Proc Natl Acad Sci USA 1987, 84: 1005-1009. 10.1073/pnas.84.4.1005PubMedCentralCrossRefPubMed
10.
go back to reference Ding XR, Yang J, Sun DC, Lou SK, Wang SQ: Whole genome expression profiling of hepatitis B virus-transfected cell line reveals the potential targets of anti-HBV drugs. Pharmacogenomics J 2008, 8: 61-70. 10.1038/sj.tpj.6500459CrossRefPubMed Ding XR, Yang J, Sun DC, Lou SK, Wang SQ: Whole genome expression profiling of hepatitis B virus-transfected cell line reveals the potential targets of anti-HBV drugs. Pharmacogenomics J 2008, 8: 61-70. 10.1038/sj.tpj.6500459CrossRefPubMed
11.
go back to reference Li GQ, Xu WZ, Wang JX, Deng WW, Li D, Gu HX: Combination of small interfering RNA and lamivudine on inhibition of human B virus replication in HepG2.2.15 cells. World J Gastroenterol 2007, 13: 2324-2327.PubMedCentralCrossRefPubMed Li GQ, Xu WZ, Wang JX, Deng WW, Li D, Gu HX: Combination of small interfering RNA and lamivudine on inhibition of human B virus replication in HepG2.2.15 cells. World J Gastroenterol 2007, 13: 2324-2327.PubMedCentralCrossRefPubMed
12.
go back to reference Xin XM, Li GQ, Guan XR, Li D, Xu WZ, Jin YY, Gu HX: Combination therapy of siRNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cell. Hepatogastroenterology 2008, 55: 2178-2183.PubMed Xin XM, Li GQ, Guan XR, Li D, Xu WZ, Jin YY, Gu HX: Combination therapy of siRNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cell. Hepatogastroenterology 2008, 55: 2178-2183.PubMed
13.
go back to reference Otsuka M, Aizaki H, Kato N, Suzuki T, Miyamura T, Omata M, Seki N: Differential cellular gene expression induced by hepatitis B and C viruses. Biochem Biophys Res Commun 2003, 300: 443-447. 10.1016/S0006-291X(02)02861-9CrossRefPubMed Otsuka M, Aizaki H, Kato N, Suzuki T, Miyamura T, Omata M, Seki N: Differential cellular gene expression induced by hepatitis B and C viruses. Biochem Biophys Res Commun 2003, 300: 443-447. 10.1016/S0006-291X(02)02861-9CrossRefPubMed
14.
go back to reference Wang J, Jiang D, Zhang H, Lv S, Rao H, Fei R, Wei L: Proteome responses to stable hepatitis B virus transfection and following interferon alpha treatment in human liver cell line HepG2. Proteomics 2009, 9: 1672-1682. 10.1002/pmic.200800621CrossRefPubMed Wang J, Jiang D, Zhang H, Lv S, Rao H, Fei R, Wei L: Proteome responses to stable hepatitis B virus transfection and following interferon alpha treatment in human liver cell line HepG2. Proteomics 2009, 9: 1672-1682. 10.1002/pmic.200800621CrossRefPubMed
15.
go back to reference Liu X, Liang J, Li G: Lipopolysaccharide promotes adhesion and invasion of hepatoma cell lines HepG2 and HepG2.2.15. Mol Biol Rep 2009, 37: 2235-2239.CrossRefPubMed Liu X, Liang J, Li G: Lipopolysaccharide promotes adhesion and invasion of hepatoma cell lines HepG2 and HepG2.2.15. Mol Biol Rep 2009, 37: 2235-2239.CrossRefPubMed
16.
go back to reference Livezey KW, Negorev D, Simon D: Hepatitis B virus-transfected Hep G2 cells demonstrate genetic alterations and de novo viral integration in cells replicating HBV. Mutat Res 2000, 452: 163-178.CrossRefPubMed Livezey KW, Negorev D, Simon D: Hepatitis B virus-transfected Hep G2 cells demonstrate genetic alterations and de novo viral integration in cells replicating HBV. Mutat Res 2000, 452: 163-178.CrossRefPubMed
17.
go back to reference Ozer A, Khaoustov VI, Mearns M, Lewis DE, Genta RM, Darlington GJ, Yoffe B: Effect of hepatocyte proliferation and cellular DNA synthesis on hepatitis B virus replication. Gastroenterology 1996, 110: 1519-1528. 10.1053/gast.1996.v110.pm8613059CrossRefPubMed Ozer A, Khaoustov VI, Mearns M, Lewis DE, Genta RM, Darlington GJ, Yoffe B: Effect of hepatocyte proliferation and cellular DNA synthesis on hepatitis B virus replication. Gastroenterology 1996, 110: 1519-1528. 10.1053/gast.1996.v110.pm8613059CrossRefPubMed
18.
go back to reference Huang YQ, Wang LW, Yan SN, Gong ZJ: Effects of cell cycle on telomerase activity and on hepatitis B virus replication in HepG2 2.2.15 cells. Hepatobiliary Pancreat Dis Int 2004, 3: 543-547.PubMed Huang YQ, Wang LW, Yan SN, Gong ZJ: Effects of cell cycle on telomerase activity and on hepatitis B virus replication in HepG2 2.2.15 cells. Hepatobiliary Pancreat Dis Int 2004, 3: 543-547.PubMed
19.
go back to reference Friedrich B, Wollersheim M, Brandenburg B, Foerste R, Will H, Hildt E: Induction of anti-proliferative mechanisms in hepatitis B virus producing cells. J Hepatol 2005, 43: 696-703. 10.1016/j.jhep.2005.02.026CrossRefPubMed Friedrich B, Wollersheim M, Brandenburg B, Foerste R, Will H, Hildt E: Induction of anti-proliferative mechanisms in hepatitis B virus producing cells. J Hepatol 2005, 43: 696-703. 10.1016/j.jhep.2005.02.026CrossRefPubMed
20.
go back to reference Tong A, Wu L, Lin Q, Lau QC, Zhao X, Li J, Chen P, Chen L, Tang H, Huang C, Wei YQ: Proteomic analysis of cellular protein alterations using a hepatitis B virus-producing cellular model. Proteomics 2008, 8: 2012-2023. 10.1002/pmic.200700849CrossRefPubMed Tong A, Wu L, Lin Q, Lau QC, Zhao X, Li J, Chen P, Chen L, Tang H, Huang C, Wei YQ: Proteomic analysis of cellular protein alterations using a hepatitis B virus-producing cellular model. Proteomics 2008, 8: 2012-2023. 10.1002/pmic.200700849CrossRefPubMed
21.
go back to reference el-Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, Pietenpol JA, Burrell M, Hill DE, Wang Y, et al.: WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 1994, 54: 1169-1174.PubMed el-Deiry WS, Harper JW, O'Connor PM, Velculescu VE, Canman CE, Jackman J, Pietenpol JA, Burrell M, Hill DE, Wang Y, et al.: WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 1994, 54: 1169-1174.PubMed
22.
go back to reference el-Deiry WS: p21/p53, cellular growth control and genomic integrity. Curr Top Microbiol Immunol 1998, 227: 121-137.PubMed el-Deiry WS: p21/p53, cellular growth control and genomic integrity. Curr Top Microbiol Immunol 1998, 227: 121-137.PubMed
23.
go back to reference Ahn JY, Jung EY, Kwun HJ, Lee CW, Sung YC, Jang KL: Dual effects of hepatitis B virus X protein on the regulation of cell-cycle control depending on the status of cellular p53. J Gen Virol 2002, 83: 2765-2772.CrossRefPubMed Ahn JY, Jung EY, Kwun HJ, Lee CW, Sung YC, Jang KL: Dual effects of hepatitis B virus X protein on the regulation of cell-cycle control depending on the status of cellular p53. J Gen Virol 2002, 83: 2765-2772.CrossRefPubMed
24.
go back to reference Masaadeh HA, Hayajneh WA, Alqudah EA: Hepatitis B virus genotypes and lamivudine resistance mutations in Jordan. World J Gastroenterol 2008, 14: 7231-7234. 10.3748/wjg.14.7231PubMedCentralCrossRefPubMed Masaadeh HA, Hayajneh WA, Alqudah EA: Hepatitis B virus genotypes and lamivudine resistance mutations in Jordan. World J Gastroenterol 2008, 14: 7231-7234. 10.3748/wjg.14.7231PubMedCentralCrossRefPubMed
25.
go back to reference Tonetto PA, Goncales NS, Fais VC, Vigani AG, Goncales ES, Feltrin A, Goncales FL Jr: Hepatitis B virus: molecular genotypes and HBeAg serological status among HBV-infected patients in the southeast of Brazil. BMC Infect Dis 2009, 9: 149. 10.1186/1471-2334-9-149PubMedCentralCrossRefPubMed Tonetto PA, Goncales NS, Fais VC, Vigani AG, Goncales ES, Feltrin A, Goncales FL Jr: Hepatitis B virus: molecular genotypes and HBeAg serological status among HBV-infected patients in the southeast of Brazil. BMC Infect Dis 2009, 9: 149. 10.1186/1471-2334-9-149PubMedCentralCrossRefPubMed
26.
go back to reference Zumbika E, Ruan B, Xu CH, Ni Q, Hou W, Chen Z, Liu KZ: HBV genotype characterization and distribution in patients with HBV-related liver diseases in Zhejiang Province, P.R. China: possible association of co-infection with disease prevalence and severity. Hepatobiliary Pancreat Dis Int 2005, 4: 535-543.PubMed Zumbika E, Ruan B, Xu CH, Ni Q, Hou W, Chen Z, Liu KZ: HBV genotype characterization and distribution in patients with HBV-related liver diseases in Zhejiang Province, P.R. China: possible association of co-infection with disease prevalence and severity. Hepatobiliary Pancreat Dis Int 2005, 4: 535-543.PubMed
27.
go back to reference Chan HL, Wong GL, Tse CH, Chim AM, Yiu KK, Chan HY, Sung JJ, Wong VW: Hepatitis B virus genotype C is associated with more severe liver fibrosis than genotype B. Clin Gastroenterol Hepatol 2009, 7: 1361-1366. 10.1016/j.cgh.2009.08.004CrossRefPubMed Chan HL, Wong GL, Tse CH, Chim AM, Yiu KK, Chan HY, Sung JJ, Wong VW: Hepatitis B virus genotype C is associated with more severe liver fibrosis than genotype B. Clin Gastroenterol Hepatol 2009, 7: 1361-1366. 10.1016/j.cgh.2009.08.004CrossRefPubMed
28.
go back to reference You J, Sriplung H, Chongsuvivatwong V, Geater A, Zhuang L, Huang JH, Chen HY, Yu L, Tang BZ: Profile, spectrum and significance of hepatitis B virus genotypes in chronic HBV-infected patients in Yunnan, China. Hepatobiliary Pancreat Dis Int 2008, 7: 271-279.PubMed You J, Sriplung H, Chongsuvivatwong V, Geater A, Zhuang L, Huang JH, Chen HY, Yu L, Tang BZ: Profile, spectrum and significance of hepatitis B virus genotypes in chronic HBV-infected patients in Yunnan, China. Hepatobiliary Pancreat Dis Int 2008, 7: 271-279.PubMed
29.
go back to reference Parkin DM, Bray F, Ferlay J, Pisani P: Global cancer statistics, 2002. CA Cancer J Clin 2005, 55: 74-108. 10.3322/canjclin.55.2.74CrossRefPubMed Parkin DM, Bray F, Ferlay J, Pisani P: Global cancer statistics, 2002. CA Cancer J Clin 2005, 55: 74-108. 10.3322/canjclin.55.2.74CrossRefPubMed
31.
go back to reference Chandra PK, Biswas A, Datta S, Banerjee A, Panigrahi R, Chakrabarti S, De BK, Chakravarty R: Subgenotypes of hepatitis B virus genotype D (D1, D2, D3 and D5) in India: differential pattern of mutations, liver injury and occult HBV infection. J Viral Hepat 2009, 16: 749-756. 10.1111/j.1365-2893.2009.01129.xCrossRefPubMed Chandra PK, Biswas A, Datta S, Banerjee A, Panigrahi R, Chakrabarti S, De BK, Chakravarty R: Subgenotypes of hepatitis B virus genotype D (D1, D2, D3 and D5) in India: differential pattern of mutations, liver injury and occult HBV infection. J Viral Hepat 2009, 16: 749-756. 10.1111/j.1365-2893.2009.01129.xCrossRefPubMed
Metadata
Title
Hepatitis B virus induces G1 phase arrest by regulating cell cycle genes in HepG2.2.15 cells
Authors
Tianzhen Wang
Ran Zhao
Yiqi Wu
Dan Kong
Lei Zhang
Di Wu
Chao Li
Chong Zhang
Zuxi Yu
Xiaoming Jin
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Virology Journal / Issue 1/2011
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/1743-422X-8-231

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