【摘要】 目的:研究大戟苷对人宫颈癌Hela细胞凋亡的诱导作用及其机制。方法:取Hela细胞分为空白对照组、顺铂组(阳性对照,10 mg/L)和大戟苷低、中、高剂量组(50、100、200 mg/L),分别加入相应药物进行培养。药物作用24、48、72 h后,采用MTT法检测细胞增殖抑制率。药物作用48 h后,采用流式细胞术检测细胞凋亡率,采用Hoechst 33258染色法检测细胞核的形态变化;采用Western blot法检测细胞色素C(Cyt-C)、Bcl-2、Bax、Caspase-3、Caspase-8、Caspase-9和Caspase-10蛋白表达水平。结果:与空白对照组比较,顺铂组和大戟苷各剂量组细胞增殖抑制率和细胞凋亡率均显著升高(P<0.05或P<0.01),细胞核浓染,或有变形、缩小、碎裂,或出现凋亡小体。与空白对照组比较,大戟苷低、中、高剂量组细胞中Cyt-C、Caspase-8和Caspase-9蛋白表达水平均显著升高,Bcl-2蛋白表达水平和Bcl-2/Bax比值均显著降低(P<0.05或P<0.01);大戟苷中、高剂量组细胞中Bax、Caspase-3和Caspase-10蛋白表达水平均显著升高(P<0.05或P<0.01)。结论:大戟苷能显著抑制Hela细胞增殖、促进细胞凋亡,其作用可能是通过激活Caspase依赖的线粒体凋亡途径来实现的。更多还原

【Abstract】 OBJECTIVE:To study induction effect of euphornin on the apoptosis of cervical cancer Hela cells and its mechanism. METHODS:The cervical cancer Hela cells were divided into blank control group,cisplatin group(positive control,10 mg/L)and euphornin low-dose,medium-dose and high-dose groups(50,100,200 mg/L). They were treated with relevant medicine. The inhibitory effect of Hela cells proliferation was tested by MTT assay after 24,48,72 h of medicine treatment. The apoptotic rate of Hela cells was measured by flow cytometry after 48 h of medicine treatment. Morphology of nucleus was detected by Hoechst 33258 staining. The protein expression of Cyt-C,Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9 and Caspase-10 were detected by Western blot assay. RESULTS:Compared with blank control group,inhibitory rate of cell proliferation and cell apoptosis rate were increased significantly in cisplatin group and euphornin groups(P<0.05 or P<0.01),and obvious staining,deformation,shrinking,fragmentation or apoptotic bodies was found in nucleus. Compared with blank control group,the protein expression levels of Cyt-C,Caspase-8 and Caspase-9 in euphornin low-dose,medium-dose and high-dose groups were increased significantly,while the protein expression level of Bcl-2 and Bcl-2/Bax ratio were decreased significantly(P<0.05 or P<0.01);the protein expression levels of Bax,Caspase-3 and Caspase-10 in euphornin medium-dose and high-dose groups were increased significantly(P<0.05 or P<0.01). CONCLUSIONS:Euphornin can significantly inhibit the proliferation of Hela cell and promote cell apoptosis,the effect of which will be achieved by activating the Caspase-dependent mitochondrion apoptosis pathway.更多还原