The mechanism of apoliprotein A1 down-regulated by Hepatitis B virus
Tóm tắt
Hepatitis B virus (HBV) infection correlated with the development of cirrhosis, liver failure and hepatocellular carcinoma (HCC), poses a huge health burden on the global community. However, the pathogenesis of chronic hepatitis B (CHB) remains unclear. Apolipoprotein A1 (ApoA1) mainly secreted by hepatocytes, represents the major protein component of high-density lipoprotein. ApoA1 secretion may be disrupted by HBV infection. In this study, we mainly investigated the molecular mechanism of ApoA1 down regulated by HBV for revealing the pathogenesis of CHB. ApoA1 expression in livers of CHB patients as well as healthy controls were performed by Real-time PCR (RT-PCR) and Western blot. The serum ApoA1 levels were measured by Enzymed-linked immunosorbent assay (ELISA). Expression of ApoA1 mRNA and protein levels were performed by RT-PCR and Western blot in human hepatoma HepG2 cells and subline HepG2.2.15 cells. HBV expression construct, pHBV1.3 were transfected into HepG2, the changes of ApoA1 mRNA and protein expression were detected by RT-PCR and Western blot. To further study the mechanism of ApoA1 down regulation by HBV, 11 CpG islands in ApoA1 promotor were tested for DNA methylation status by MSP. HepG2.2.15 cell lines were treated with DNA methyltransferase inhibitor 5-aza-deoxycytidine (5-aza-dC), then, expression of ApoA1 mRNA and HBV particles in the supernatant, as well as ApoA1 protein levels were detected by RT-PCR and Western blot. Secretion of HBsAg and HBeAg in HepG2 cells cotransfected with pApoA1 and pHBV1.3 constructs was tested by ELISA. Meanwhile, secretion of HBsAg and HBeAg in the supernatant were quantified by ELISA in the HepG2.2.15 cells treated with 5-aza-dC plus ApoA1 siRNA. Expression of ApoA1 mRNA and protein levels, as well as serum ApoA1 levels in CHB patients were decreased corresponding healthy controls in vivo. In addition, the expression of ApoA1 mRNA and protein levels were down regulated in HepG2.2.15 cells correponding HepG2 cells, 11 CpG islands in ApoA1 promoter were tested for methylation status by MSP in HepG2.2.15 cells compared to HepG2 cells, while two CpG islands were found hypermethylated. Expression of ApoA1 mRNA and protein levels were increased in HepG2.2.15 cells treated with DNA methyltransferase inhibitor 5-aza-dC. Furthermore, overexpression of ApoA1 can enhance HBV expression in HepG2 cells while the inhibitory effect of 5-aza-dC on HBV expression was completely abolished by blocking 5-aza-dC-induced up-regulation of ApoA1 using RNAi. Epigenetic silencing of ApoA1 gene expression by CpG island DNA hypermethylation induced by HBV may contribute to the pathogenesis of CHB.
Tài liệu tham khảo
Jiang W, Zheng L, Yang Q, Huang Z, Wang X. Investigation into the effect of hepatitis B virus on apoliprotein A1 expression and its mechanism. Lipids Health Dis. 2014;13:130.
Hao J, Jin W, Li X, Wang S, Zhang X, Fan H, Li C, Chen L, Gao B, Liu G, Meng S. Inhibition of alpha interferon (IFN-alpha)-induced microRNA-122 negatively affects the anti-hepatitis B virus efficiency of IFN-alpha. J Virol. 2013;87:137–47.
Teng YC, Shen ZQ, Kao CH, Tsai TF. Hepatocellular carcinoma mouse models: Hepatitis B virus-associated hepatocarcinogenesis and haploinsufficient tumor suppressor genes. World J Gastroenterol. 2016;22:300–25.
Huang LH, Elvington A, Randolph GJ. The role of the lymphatic system in cholesterol transport. Front Pharmacol. 2015;6:182.
Park SW, Lee EH, Lee EJ, Kim HJ, Bae DJ, Han S, Kim D, Jang AS, Uh ST, Kim YH, et al. Apolipoprotein A1 potentiates lipoxin A4 synthesis and recovery of allergen-induced disrupted tight junctions in the airway epithelium. Clin Exp Allergy. 2013;43:914–27.
Morin EE, Guo L, Schwendeman A, Li XA. HDL in sepsis - risk factor and therapeutic approach. Front Pharmacol. 2015;6:244.
Aluganti Narasimhulu C, Selvarajan K, Brown M, Parthasarathy S. Cationic peptides neutralize Ox-LDL, prevent its uptake by macrophages, and attenuate inflammatory response. Atherosclerosis. 2014;236:133–41.
Toptas B, Gormus U, Ergen A, Gurkan H, Kelesoglu F, Darendeliler F, Bas F, Dalan AB, Izbirak G, Isbir T. Comparison of lipid profiles with APOA1 MspI polymorphism in obese children with hyperlipidemia. In Vivo. 2011;25:425–30.
Mohamadkhani A, Jazii FR, Sayehmiri K, Jafari-Nejad S, Montaser-Kouhsari L, Poustchi H, Montazeri G. Plasma myeloperoxidase activity and apolipoprotein A-1 expression in chronic hepatitis B patients. Arch Iran Med. 2011;14:254–8.
Zhu C, Zhang R, Liu L, Rasool ST, Mu Y, Sun W, Hao Q, Liu F, Zhu Y, Wu J. Hepatitis B virus enhances interleukin-27 expression both in vivo and in vitro. Clin Immunol. 2009;131:92–7.
Norton PA, Gong Q, Mehta AS, Lu X, Block TM. Hepatitis B virus-mediated changes of apolipoprotein mRNA abundance in cultured hepatoma cells. J Virol. 2003;77:5503–6.
Zhang T, Xie N, He W, Liu R, Lei Y, Chen Y, Tang H, Liu B, Huang C, Wei Y. An integrated proteomics and bioinformatics analyses of hepatitis B virus X interacting proteins and identification of a novel interactor apoA-I. J Proteomics. 2013;84:92–105.
Zhang J, Fu LL, Tian M, Liu HQ, Li JJ, Li Y, He J, Huang J, Ouyang L, Gao HY, Wang JH. Design and synthesis of a novel candidate compound NTI-007 targeting sodium taurocholate cotransporting polypeptide [NTCP]-APOA1-HBx-Beclin1-mediated autophagic pathway in HBV therapy. Bioorg Med Chem. 2015;23:976–84.
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 U S A. 1987;84:1005–9.
Tsezou A, Iliopoulos D, Malizos KN, Simopoulou T. Impaired expression of genes regulating cholesterol efflux in human osteoarthritic chondrocytes. J Orthop Res. 2010;28:1033–9.
Zhong S, Tang MW, Yeo W, Liu C, Lo YM, Johnson PJ. Silencing of GSTP1 gene by CpG island DNA hypermethylation in HBV-associated hepatocellular carcinomas. Clin Cancer Res. 2002;8:1087–92.
Cheong JY, Kim DJ, Hwang SG, Yang JM, Kim YB, Park YN, Cho SW. Serum markers for necroinflammatory activity in patients with chronic viral hepatitis and normal or mildly elevated aminotransferase levels. Liver Int. 2011;31:1352–8.
Lee HJ, Seo YS, Kim DJ, Kang HS, An H, Kim JH, Cheong JY, Yim HJ, Yeon JE, Lee HS, et al. Application of the HALF index obviates the need for liver biopsy in half of all patients with chronic hepatitis B. J Gastroenterol Hepatol. 2011;26:987–95.
Stenzig J, Hirt MN, Loser A, Bartholdt LM, Hensel JT, Werner TR, Riemenschneider M, Indenbirken D, Guenther T, Muller C, et al. DNA methylation in an engineered heart tissue model of cardiac hypertrophy: common signatures and effects of DNA methylation inhibitors. Basic Res Cardiol. 2016;111:9.
Menschikowski M, Hagelgans A, Nacke B, Jandeck C, Sukocheva O, Siegert G. Epigenetic control of phospholipase A2 receptor expression in mammary cancer cells. BMC Cancer. 2015;15:971.
Li H, Yang F, Gao B, Yu Z, Liu X, Xie F, Zhang J. Hepatitis B virus infection in hepatocellular carcinoma tissues upregulates expression of DNA methyltransferases. Int J Clin Exp Med. 2015;8:4175–85.
Zhou XQ, Huang SY, Zhang DS, Zhang SZ, Li WG, Chen ZW, Wu HW. Effects of 5-aza-2'deoxycytidine on RECK gene expression and tumor invasion in salivary adenoid cystic carcinoma. Braz J Med Biol Res. 2015;48:254–60.