PLNCX-TNFα基因转染对人胆管癌细胞生长的影响

EFFECTS OF RECOMBINANT REVERSE TRANSCRIPT VIRUS TNFα GENE TRANSDUCTION ON PROLIFERATION AND APOPTOSIS OF HUMAN CHOLANGIOCARCINOMA CELLS

目的 研究 L NCX- TNFα基因过表达对人胆管癌细胞的作用。方法 构建 L NCX- TNFα复合体 ,培养人胆管癌细胞 ,L NCX- TNFα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 重组体 L NCX- TNFα基因转染人胆管癌细胞后 ,能抑制细胞的生长和集落形成 ;流式细胞技术证实 ,L NCX- TNFα能诱导细胞发生凋亡并导致其发生 G1 期阻滞 ,G2 - M期和 S期比例明显下降 ,凋亡细胞数也明显增加。结论 TNFα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 TNFα 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 .The transfected cell were 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 TNFα gene,cells growth rate were obviously inhibited and resulted in an accumlation of cells 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 phase obviously decreased from 18.32% to 10.34% and from 52.03% to 2.02%,respectively,as compared with group.Colony formation in these LNCX-TNFα-infected cells greatly decreased compared with LNCX-TNFα-uninfected cells.The apoptotic index was increased in LNCX-TNF α infected cells from 5.83% to 34.76%.Conclusion Tranfer of TNFα gene could inhibit the growth of human cholangiocarcinoma cells and induce cell apoptosis.