目的观察丙型肝炎病毒核心蛋白(HCVc)阳性肝内胆管癌细胞中人端粒酶逆转录酶(hTERT)的表达,并探讨DNA甲基转移酶1(DNMTl)对hTERT表达的影响。方法将HCVc质粒瞬转入肝内胆管癌细胞株RBE,5-氮杂-2’-脱氧胞嘧啶核苷(AZA)处理转染后细胞;Westernblot法及逆转录.聚合酶链反应(RT.PCR)检测细胞中hTERT及DNMTl的mRNA及蛋白表达。结果比较空白对照组,转染HCVc质粒后,RBE细胞株中hTERT及DNMTl的mRNA、蛋白水平分别上调3.457±0.081、1.684±0.020、1.826±0.060、2.513±0.119(P〈0.05);AZA处理转染后细胞可逆转hTERT的升高,mRNA和蛋白水平分别下调2.755±0.098、1.491±0.045(P〈0.05)。结论DNMTl参与HCVc诱导的hTERT表达上调过程。
Objective To explore the expression of human telomerase reverse transcfiptase (hTERT) in intrahepatie cholangiocarcinoma cell line RBE transfected with hepatitis C virus core protein (HCVc) plasmid, and involvement of DNA methy ltransferase 1 ( DNMT1 ) in the up-regulation of hTERT induced by HCVc. Methods Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to analyze the expression of hTERT and DNMT1 after the transfeetion of HCVe plasmid into RBE cells. 5-Aza-2'-deoxycytidin (AZA) was used to inhibit DNMT1. Results hTERT expression level was higher in RBE cells after HCVe transfeetion and DNMT1 expression was up-regulated simultaneously. The mRNA and protein levels of hTERT were up-regulated by 3.457 ~ 0. 081 and 1. 684 ± 0. 020 respectively ( P 〈 0.05 ). And the expression levels of DNMT1 were up-regulated by 1. 826 ± 0. 060 and 2.513± 0. 11.9 respectively (P〈 0.05). After treatment with AZA, the mRNA and protein levels of hTERT were down-regulated by 2. 755 ± 0. 098 and 1. 491 ± 0. 045 respectively ( P 〈 0.05 ). Conclusion DNMT1 may take part in the up-regulation of hTERT induced by HCVc. '