摘要
目的探讨金粉蕨素(ONY)体外诱导人宫颈癌HeLa细胞凋亡的作用及其分子机制。方法体外培养人宫颈癌HeLa、CaSki、SiHa、ME180细胞,采用MTT法检测ONY对人宫颈癌HeLa、CaSki、SiHa、ME180细胞生长抑制率的影响;Hoechst33258染色检测ONY对人宫颈癌HeLa细胞凋亡的形态学变化;流式细胞术(FCM)检测ONY对HeLa细胞凋亡率的影响;DNA琼脂糖电泳检测HeLa细胞凋亡的DNA条带;Western blot分析凋亡相关蛋白表达变化。结果 ONY对人宫颈癌HeLa、CaSki、SiHa、ME180细胞生长有较强的抑制作用,呈剂量和时间依赖性,以HeLa细胞对ONY最为敏感,作用24 h的IC50值为10.48μg·mL-1。经ONY作用于HeLa细胞24 h后,细胞出现典型的凋亡形态学改变,表现出典型的凋亡特征的亚二倍体峰,且呈剂量依赖性,同时出现典型的凋亡DNA条带;同时,cytochrome c、caspase-9和caspase-3蛋白表达增加,bcl-2蛋白表达下调,而bax蛋白表达不变,Bcl-2/Bax比值下调(P<0.05)。结论 ONY可能通过调控线粒体途径相关蛋白抑制宫颈癌HeLa细胞增殖并诱导其凋亡。
Objective To investigate the effects and mechanisms of Onychin （ONY） on the apoptosis in human cervical can-cer HeLa cell line in vitro. Methods Human cervical cancer lines HeLa, CaSki, SiHa, ME180 were cultured in vitro. The effects of ONY on the growth inhibition ratio of HeLa, CaSki, SiHa, ME180 cells were evaluated by the MTT assay. The effects of ONY on the apoptosis of HeLa cells were detected through morphological observation by Hoechst33258 staining, flow cytometric analysis （FCM）, and agarose gel electrophoresis. The expressions of proteins related to apoptosis were analyzed by western blot. Results ONY significantly inhibited the growth and proliferation of human cervical cancer cell lines HeLa, CaSki, SiHa, ME180 in a dose-and time-dependent manner. HeLa cell was the most sensitive one among the four kinds of cells, and its IC50 was 10.48μg·mL-1 for 24 h. The cells treated with ONY for 24 h showed typical morphological change of apoptosis, increased cells of sub-G1 population in dose-dependent manner and typical DNA ladder of agarose gel electrophoresis. Western blot showed the expressions of cytochrome-c, caspase-9 and caspase-3 protein were up-regulated, and protein expression of Bcl-2 was depressed, while Bax protein expression level had not changed. The ratio of Bcl-2/Bax was down-regulated after treatment with ONY in a concentration dependent manner. Conclusion ONY may inhibit the proliferation and induce the apoptosis of human cervical cancer HeLa cells through mitochondrial apoptosis pathway in vitro.
出处
《肿瘤药学》
CAS
2014年第3期176-181,共6页
Anti-Tumor Pharmacy
关键词
宫颈癌
金粉蕨素
凋亡
增殖
线粒体凋亡途径
Cervical cancer
Onychin
Apoptosis
Proliferation
Mitochondrial apoptosis pathway
