Nutlins类似物NL-86诱导HeLa细胞凋亡的研究

目的 观察新型Nutlins类似物NL-86在体外诱导宫颈癌HeLa细胞凋亡的作用,并初步探讨其作用的分子机制.方法 采用MTT法检测NL-86化合物对HeLa细胞增殖的影响;用 FITC-Annexin V及碘化丙锭（PI）双染法,通过流式细胞仪（FCM）检测NL-86诱导HeLa细胞凋亡情况;用Western 印迹检测PARPpoly（ADP-ribose）polymerase]、pro-caspase 3、8、9的变化,并利用活力检测试剂盒检测caspase 3、8、9的活力,初步确定其诱导凋亡的通路.结果 不同浓度的NL-86 对于HeLa细胞的存活率均具有一定影响,并以浓度依赖的方式诱导HeLa细胞凋亡;随着NL-86浓度的增加,PARP被切割、HeLa 细胞pro-casepase 3、8的含量下降,但 pro-caspase 9无变化.活力测定结果显示caspase 3、8被激活,caspase 9无变化.结论 NL-86化合物能够有效地在体外诱导HeLa 细胞凋亡,且可能依赖于caspase 3、8相关的死亡受体凋亡途径,为进一步研发治疗宫颈癌等肿瘤疾病的药物奠定了实验基础.

Apoptotic Induction of HeLa Cells by A New Nutlins Analog NL-86 Compound

Objective To observe the apoptosis of HeLa cells induced in vitro by nutlins analog NL-86 and investigate the relevant molecular mechanism. Methods The effect of NL-86 on proliferation of HeLa cells was determined by MTT method. The apoptosis of HeLa cells were determined by flow cytometry with FITC-Annexin V and PI. The activity of caspase 3/8/9 were determined by caspase activity assay kit and western blot. Results NL-86 conferred inhibitory effect on proliferation and induced apoptosis of HeLa cells in dose-dependent manner. The activity of caspase 3/8 in HeLa cells increased with increase of concentration of NL-86, while caspase 9 showed no changes. Conclusion NL-86 induced apoptosis of HeLd cells in vitro through caspase 3/8 activity and death receptor-mediated signal pathway .