金丝桃苷对宫颈癌Hela细胞凋亡及抗氧化能力的影响

EFFECTS OF HYPEROSIDE ON APOPTOSIS AND ANTIOXIDATIVE CAPACITY OF CERVICAL CANCER HELA CELLS

目的探索金丝桃苷对宫颈癌Hela细胞凋亡、抗氧化能力的影响。方法采用不同浓度的金丝桃苷(0、25、50、100、200μmol/L)处理体外培养Hela细胞12 h、24 h、48 h,噻唑蓝(MTT)法检测Hela细胞各浓度不同时间点细胞增殖情况以筛选出适宜的金丝桃苷浓度与作用时间点;giemsa染色观察比较观察组(金丝桃苷:100μmol/L)与对照组(空白对照)Hela细胞24 h形态变化;流式细胞仪检测Hela细胞凋亡情况;免疫组化检测caspase-3与caspase-8蛋白表达;ELISA法检测Hela细胞MDA、谷胱甘肽(GSH)的水平,SOD、过氧化氢酶(CAT)活力;qPCR检测Hela细胞VEGF、Bcl-2、Bax、p53表达水平。结果随着金丝桃苷作用时间的延长,Hela细胞存活率逐渐降低并且存在剂量依赖效应;观察组Hela细胞数目减少、光泽度降低、细胞体积变小且形状发生改变;观察组Hela细胞凋亡率明显高于对照组;观察组Hela细胞caspase-3与caspase-8蛋白表达量高于对照组;观察组Hela细胞MDA含量高于对照组,而GSH水平显著低于对照组;观察组Hela细胞SOD、CAT活力显著低于对照组;观察组Hela细胞VEGF、Bcl-2表达量降低,而Bax、p53表达水平高于对照组。结论金丝桃苷可以通过调控凋亡相关基因和蛋白的表达、促进细胞凋亡、降低抗氧化能力,从而抑制宫颈癌Hela细胞生长。

Objective To explore the effects of Hyperoside on apoptosis and antioxidant capacity of cervical cancer Hela cells.Methods Hela cells were cultured in vitro for 12 h,24 h and 48 h by treatment of different concentrations of Hyperoside(0,25,50,100,200μmol/L).Methyl thiazolyl tetrazolium(MTT)assay was used to detect the proliferation of Hela cells at different concentrations and time points to select appropriate hyperoside concentration and time point.Giemsa staining was used to observe and compare the 24 h morphological changes of Hela cells between observation group(hyperside:100μmol/L)and control group(blank control).The apoptosis of Hela cells was detected by flow cytometry.The protein expression levels of caspase-3 and caspase-8 were detected by immunohistochemistry.The levels of MDA and glutathione(GSH)and activities of SOD and catalase(CAT)of Hela cells were detected by ELISA.The expression levels of VEGF,Bcl-2,Bax and p53 in Hela cells were detected by qPCR.Results With the prolongation of action of hyperoside,the survival rate of Hela cells WAS gradually decreased and showed dose-dependent effects.The number of Hela cells was decreased,the glossiness was decreased and the cell volume became smaller and the shape changed in observation group.The apoptosis rate of Hela cells in observation group was significantly higher than that in control group.The protein expression levels of caspase-3 and caspase-8 of Hela cells in observation group were higher than those in control group.The MDA content of Hela cells in observation group was higher than that in control group while the GSH level was significantly lower than that in control group.Theactivities of SOD and CAT of Hela cells in observation group were significantly lower than those in control group.The expression levels of VEGF and Bcl-2 in Hela cells were decreased while the expression levels of Bax and p53 were higher than those in control group.Conclusion Hyperoside can inhibit the growth of cervical cancer Hela cells by regulating the expressions of apoptosis-related genes and proteins,promoting apoptosis and reducing antioxidant capacity.