肝细胞癌中抑制细胞凋亡相关基因的差异表达

Comparative analysis of cell apoptosis inhibitory genes involved in hepatoma

目的:研究肝癌细胞系的基因表达概况,筛选表达差异的抑制细胞凋亡相关基因,探索肝细胞癌发生发展机制及肝细胞癌治疗的新靶点。方法:通过基因芯片技术检测肝癌细胞系Bel-7402、Hep-3B与胎儿肝细胞的全基因组序列,筛选表达差异的抑制细胞凋亡相关基因,采用RT-PCR、免疫组化对部分差异表达基因进行了验证。结果:在检测的54614个探针组中,Bel-7402、Hep-3B两株肝癌细胞与胎儿肝脏细胞相比表达共同上调2倍及2倍以上的有1452个基因;表达下调2倍及2倍以上的有2054个基因。与抑制细胞凋亡相关表达上调的基因有12个,选取2个基因进行RT-PCR、免疫组化验证,结果与基因芯片结论基本一致。结论:肝细胞癌中有众多的抑制细胞凋亡基因表达增高,通过影响TNF通路、bcl-2蛋白等多种方式共同抑制细胞凋亡,促进肿瘤细胞增殖。对这些基因的研究有助于阐明肝细胞癌发生发展机制,为肝细胞癌的治疗提供新的靶点。

Objective：To explore the profiles of hepatoma cell lines-specific gene expression and identify the differential expressed cell apoptosis inhibitory genes for exploring hepatocarcinogenesis and new therapeutic targets. Methods：The profiles of gene expression of hepatoma cell lines Bel-7402, Hep-3B and fetal liver cells were analyzed by gene chips. The differential expressed cell apoptosis inhibitory genes were screened. The selected candidate genes were confirmed by RT-PCR in three kinds of cell lines and by immuno-histochemistry in 20 specimens of HCC and adjacent noncancerous tissues. Results：Of the detected 54 614 probe sets,compared with the fetal liver cells, 1 452 genes were up-regulated by more than 2 folds and 2 054 genes were down-regulated by more than 2 folds in the hepatoma cell lines Bel-7402 and Hep-3B,in which 12 differential expressed cell apoptosis inhibitory genes were screened. The results of RT-PCR showed that the expressions of SODD and Survivin in hepatoma cell lines were higher than those in fetal liver cells （P 〈 0.05 ）. The positive rates of SODD and Survivin were significantly higher in HCC than in adjacent noncancerous tissues （65% vs. 15%, and 75% vs. 10%,P 〈 0.05）. Conclusion：Many cell apoptosis inhibitory genes were up-regulated in HCC,which can inhibit cancer cell apoptosis and promote cancer cel] proliferation by effecting TNF pathway,bcl-2 protein expression and so on. Analysis of these genes can provide important clues for disclosing the mechanisms of hepatocellular carcinoma pathogenesis and supplying possible new therapeutic targets.