氧化苦参碱对人宫颈癌SiHa细胞株增殖及凋亡的影响

目的研究氧化苦参碱对体外人宫颈癌SiHa细胞株增殖活性及凋亡的影响。方法对人宫颈癌SiHa细胞株进行体外培养,经氧化苦参碱处理的细胞组为实验组,未经处理组为对照组。采用MTT细胞存活实验检测氧化苦参碱对入宫颈鳞癌SiHa细胞的增殖影响,并计算IC50;倒置相差显微镜下观察不同浓度的氧化苦参碱作用48h后SiHa细胞的形态改变;Hoechst33258染色法和Western blot法检测氧化苦参碱对SiHa细胞核及凋亡相关蛋白(P53、Bax及Bcl-2)表达水平的影响。结果 MTT结果显示氧化苦参碱剂量-时间依耐性抑制人宫颈癌SiHa细胞的体外增殖(P0.05),计算24 h、48 h和72 h的IC50分别为(1 028.41±3.57)μg/ml、(701.72±6.01)μg/ml和(406.88±2.15)μg/ml;氧化苦参碱处理48 h后,SiHa细胞的形态特征及数目发生显著变化,且随氧化苦参碱浓度的增加而愈加明显;Hoechst33258染色证实氧化苦参碱处理后SiHa细胞发生凋亡,可见典型的凋亡特征;Western blot结果证实氧化苦参碱(700μg/ml、1 600μg/ml)处理48 h后,和对照组比较,药物处理组细胞凋亡周期蛋白P53、Bax表达上调(P0.05),而Bcl-2表达下调(P0.05),Bax/Bcl-2的比率明显增加(P0.05)。结论氧化苦参碱可抑制体外人宫颈癌SiHa细胞的体外增殖活性发挥其抗肿瘤作用,其作用机制可能诱导SiHa细胞的凋亡有关。

Effect of Oxymatrine on Cell Proliferation and Apoptosis of Human Cervical Cancer Cell Line SiHa

Objective To evaluate the proliferative effect and induction of apoptosis by oxymatrine in human cervical cancer cell line SiHa in vitro.Methods SiHa were cultured in vitro.The experimental groups were treated with oxymatrine,the oth-ers were in the control groups.Proliferation rates and IC50 values were assessed by methyl thiazolyl tetrazolium （MTT）.In-verted phase contrast microscope was used to observed the morphological alterations of SiHa human cervical cancer cells at five concentrations of oxymatrine at 48 h.The morphological changes accompanying cell apoptosis were observed using an inverted microscope after Hoechst 33258 staining.In addition,expression changes of apoptosis-related proteins involved in the P53,Bax and Bcl-2 were determined by western blotting.Results The results determined by MTT assay showed that the viability of human cervical cancer SiHa cells with oxymatrine was markedly attenuated in a concentration and time depend-ent manner（P〈0.05）.The IC50 values were calculated as （1 028.41 ±3.57）μg/ml,（701.72 ±6.01）μg/ml and （406.88 ±2.15）μg/ml at 24 h,48 h and 72 h.The morphological alterations of SiHa human cervical cancer cells changed significantly with the increase of the concentration of oxymatrine at 48 h.Also the oxymatrine-treated SiHa human cervical cancer cells displayed typical apoptosis morphological changes by the Hoechst 33258 staining at 48h.Compared with the control groups,the experimental groups resulted in remarkable increase of both P53 and Bax（P〈0.05 ）,but significantly decrease of Bcl-2（P〈0.05）in Western blot.Conclusion These results demonstrate that oxymatrine exerts antitumor effects on SiHa cells through its anti-proliferative role,and suggest the mechanism for the effect may be related to its induction of apoptosis effects.