摘要
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. ＇
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第11期2113-2116,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30872485）
