Cpd5对人卵巢癌SKOV3细胞增殖抑制和凋亡诱导作用

Proliferation inhibition and apoptosis induction in human ovarian cancer SKOV3 cells by Cpd5

目的:观察Cpd5[2-(2-巯基乙醇)-3-甲基-1,4-萘醌,Compound5]对人卵巢癌SKOV3细胞增殖和凋亡的影响。方法:MTT法观察Cpd5对SK-OV3细胞的生长抑制作用,AnnexinV/PI双标记流式细胞术检测Cpd5对SKOV3细胞的凋亡诱导,Hoechst-33258染色荧光显微镜观察细胞凋亡形态。结果:5、10、20、30、40、50、60μmol/LCpd5处理SKOV3细胞48h,生长抑制率分别为2.77%、5.19%、10.61%、41.15%、71.37%、82.90%、89.81%。不同时间点(12、24、48和72h)检测,细胞生长抑制率存在剂量-时间依赖关系。流式细胞术检测,30μmol/LCpd5处理12、24和48h后,细胞凋亡率分别达9.25%、20.07%、56.16%;50μmol/L组的细胞凋亡率明显高于30μmol/L,且随时间增加而显著增加。用40、50和60μmol/LCpd5处理细胞12h,光镜观察即可见明显的形态学改变。荧光显微镜观察:AnnexinV-EGFP/PI双染显示,实验组比阴性对照组细胞凋亡率明显增多;Cpd5作用24、48h后,Hoechst-33258染色可见细胞出现明显的凋亡形态,20μmol/L浓度组凋亡率增加,30、40、50、60μmol/L各组细胞凋亡率显著增加。结论:Cpd5能以剂量-时间依赖的方式抑制SKOV3细胞增殖并诱导凋亡,是潜在的抗卵巢癌新化合物。

AIM：To investigate the effect of Cpd5 on the proliferation inhibition and apoptosis induction in human ovarian cancer SKOV3 cells.METHODS：The growth inhibition of Cpd5 in SKOV3 cells was detected by MTT assay.AnnexinV/PI staining was employed for quantifying apoptotic cells by fluorescence microscopy and flow cytometry.The apoptotic morphology was observed by Hoechst-33258 staining.RESULTS：After Cpd5 treatment at the concentrations of 5,10,20,30,40,50 and 60 μmol/L for 48 h,the ratios of the cells growth inhibition were 2.77%,5.19%,10.61%,41.15%,71.37%,82.90% and 89.81%,respectively.The proliferation was inhibited after Cpd5 treatment at different times of 12,24,48 and 72 h,the cells were inhibited by Cpd5,in a dose-time-dependent manner.The apoptotic cells accounted for 9.25%,20.07% and 56.16%,after Cpd5 treatment by 30 μmol/L for 12,24 and 48 h,respectively.The ratio of apoptotic cells in 50 μmol/L group was significantly higher than that in 30 μmol/L group.The morphological changes were emerged after Cpd5 treatment at the concent rations of 40,50 and 60 μmol/L for 12 h.Compared with the control group,the ratios of the apoptotic cells were increased significantly in the groups induced by Cpd5 at different concentrations.The apoptotic cells were increased in the groups of Cpd5 treatment at 20 μmol/L for 24 h and 48 h by Hoechst-33258 staining.Furthermore,the ratios of apoptotic cells were increased significantly in the groups of Cpd5 treatment at 30,40,50 and 60 μmol/L Cpd5 at different time-points.CONCLUSION：Cpd5 can induce proliferation inhibition and apoptosis in SKOV3 cells,in a dose-time-dependent manner.Our data reveal that Cpd5 is a novel anti-ovarian cancer compound.