肝癌细胞系中DNMTs表达的检测及意义

Detection and Significance of Expression of DNMTs in Hepatoma Carcinoma Cell Lines

目的:检测HBV相关肝癌细胞系(Hep3B)与非HBV相关肝癌细胞系(HepG2)中甲基化转移酶(DNAmethyltransferase,DNMT)的表达,分析HBV感染与DNMT表达之间的关系。方法:应用荧光定量PCR(Flurescencequantitative-PCR,FQ-PCR)检测两种肝癌细胞系中DNMT1、DNMT2、DNMT3A、DNMT3B、GAPDH基因mRNA表达水平,并将DNMT1、DNMT2、DNMT3A和DNMT3B表达水平与参比基因进行比较。结果:DNMT1、DNMT2、DNMT3A和DNMT3B在HBV相关细胞系Hep3B中表达分别为:1.26±0.15、0.76±0.15、1.26±0.20、0.66±0.13;在非HBV相关肝癌细胞系HepG2中的表达分别为:0.64±0.11、0.62±0.12、0.48±0.12、0.36±0.10。Hep3B细胞系中DNMT1、DNMT3A和DNMT3B的表达均高于HepG2(P均<0.01)。结论:⑴HBV相关肝癌细胞系Hep3B中存在DNMT过表达;⑵HBV感染可能刺激DNMT1、DNMT3A和DNMT3B的表达,进而促进抑癌基因甲基化。

Objective To detect DNA methyltransferases(DNMTs) expression in hepatoma carcinoma cell lines(Hep3B and HepG2) and to explore the relationship between HBV infection and DNMTs expression.Methods The mRNA expression of four types DNMTs(DNMT1,DNMT2,DNMT3A,DNMT3B) was detected with FQ-PCR in Hep3B and HepG2,and then compared with the expression of referential gene.Results The mRNA expression of DNMT1,DNMT2,DNMT3A,DNMT3B was 1.26±0.15,0.76±0.15,1.26±0.20,0.66±0.13 in Hep3B and 0.64±0.11,0.62±0.12,0.48±0.12,0.36±0.10 in HepG2,respectively.The expression of DNMT1,DNMT3A,DNMT3B in Hep3B was obviously higher than those in HepG2(all P<0.01).Conclusion ⑴Over-expression of DNMTs mRNA was observed in HBV-associated cells(Hep3B);⑵HBV infection may promote the expression of DNMT1,DNMT3A and DNMT3B,and potentiate the methylation of tumor suppressor genes.