重组改构人肿瘤坏死因子联合5-氟尿嘧啶对胃癌细胞生长的抑制作用

Inhibiting effects of recombinant mutant human tumor necrosis factor and fluorouracil on BGC-823 cells

目的:研究重组改构人肿瘤坏死因子(rmhTNF)单用及与5-氟尿嘧啶(5-FU)联合应用对人胃癌BGC-823细胞系生长的抑制作用及途径。方法:细胞培养24h后随机分为4组(A组为空白对照组,B组为rmhTNF组,C组为5-FU组,D组为rmhTNF+5-FU组),加入相应药物后继续培养72h,应用TUNEL法检测4组细胞的凋亡率,免疫细胞化学法检测4组细胞中细胞色素C(Cyt-C)的表达,流式细胞仪检测4组细胞的周期分布和凋亡率。结果:①TUNEL结果显示联合用药组细胞凋亡率显著高于其他各组,各药物处理组显著高于对照组(P<0.05)。②免疫细胞化学法结果显示联合用药组细胞胞浆Cyt-C释放显著高于其他各组,各药物处理组显著高于对照组(P<0.05)。③流式细胞仪结果显示联合用药组S期细胞明显减少,凋亡率增加(P<0.05)。结论:rmhTNF可诱导BGC-823细胞凋亡,增加5-FU对细胞的杀伤作用,其作用机制与细胞凋亡的线粒体途径有关。

Aim： To observe the inhibiting effects of recombinant mutant human tumor necrosis factor（rmhTNF） and fluorouracil（5-FU） on BGC-823 cells. Methods ： BGC-823 cells were cultured for 24 h,then randomly allocated into 4 groups. Group A was control group,group B was rmhTNF group,group C was 5-FU group, and group D was rmhTNF ＋ 5-FU group, and they were exposed to the reagents for 72 h. The apoptotic cells were detected by TUNEL assay, the expression of Cyt-C protein was detected by immunocytochemistry assay,and the percentage of apoptotic cells and cell cycle distribution were de- tected by the flow cytometer. Results：The cell apoptotic rate in rmhTNF ＋ 5-FU group was significantly higher than that of either single drug group,and the apoptotic rate in medicine group was significantly higher than that of control goup（ P 〈 0.05）. Cyt-C in rmhTNF ＋ 5-FU group was higher than that of either single drug group,and the Cyt-C in medicine group was higher than that of control group（P 〈 0.05）. Arrest and apoptosis rate of cell in rmhTNF ＋ 5-FU group was significantly stronger than that of either single drug group（ P 〈 0.05）. Conclusion： rmhTNF alone could induce BGC-823 cells apoptosis. Combination of rmhTNF and 5-FU has synergetic anticancer effect on BGC-823 cells by mitochondrial pathway.