肿瘤坏死因子基因转染对人胆管癌细胞生长的影响

Effects of recombinant reverse transcript virus tumor necrosis factor alpha gene transduction on proliferation and apoptosis of human cholangiocarcinoma cells

目的 研究肿瘤坏死因子 α(TNF α)基因过表达对人胆管癌细胞生长的影响。方法构建逆转录病毒 TNF α(pLNCX TNF α)复合体 ,转染人胆管癌细胞得到稳定表达 ,通过聚合酶链式反应 (PCR)、流式细胞仪 (FCM )、免疫组织化学等方法检测基因表达、细胞生长增殖和细胞凋亡等情况。结果 pLNCX TNF α基因转染人胆管癌细胞后 ,pLNCX空质粒转染组、pLNCX TNF α基因转染组的细胞均能扩增出 70 8bp的基因片断 ,而未转染组则无 ;pLNCX TNF α基因转染组的细胞克隆抑制率为 64 % ,与对照组和 pLNCX空质粒转染组相比 ,差异性非常显著 (P <0 .0 1) ;转染的人胆管癌细胞G1期比例从 2 9.0 7%增加到 5 4.2 4% ,G2 M期和S期比例分别从 5 2 .0 3 %下降到2 .0 2 %、18.3 2 %下降到 10 .3 4% ;凋亡细胞数从 5 .83 %增加到 3 4.76%。结论 TNF α基因通过诱导肿瘤细胞发生凋亡并导致其发生G1期阻滞在肿瘤基因治疗方面发挥作用。

Objective To evaluate the inhibitory effect of tumor necrosis factor alpha (TNF-α) on proliferation and apoptosis of human cholangiocarcinoma cells.Methods Recombinant reverse transcript virus TNF-α gene was transfected into the cells of human cholangiocarcinoma in vitro,which stably overexpressed TNF-α gene.The expression of the gene in the transfected cells was detected by PCR and immunocytochemical methods respectively.The cycle pattern and apoptosis were assayed by using flow cytometry.Results After transfection of recombinant reverse transcript virus TNF-α gene,the cells growth rate were obviously inhibited and the cells arrested in G_1 phase,presenting that the proportion of the cells in G_1 phase was obviously increased from 29.07% to 54.24%,and that of the cells in S and G_2-M phases obviously decreased from 10.34% to 18.32% and from 52.03% to 2.02%,respectively,as compared with control group.Colony formation in these recombinant reverse transcript virus TNF-α-infected cells was greatly decreased as compared to the recombinant reverse transcript virus TNF-α-uninfected cells.The apoptotic index was increased in the recombinant reverse transcript virus TNF-α-infected cells from 5.83% to 34.76%.Conclusion Tranfer of TNF-α gene could inhibit the growth of human cholangiocarcinoma cells and induce cell apoptosis.