摘要
目的探讨自噬在蝙蝠葛碱诱导宫颈癌Hela细胞凋亡中的作用方法采用MTT法检测蝙蝠葛碱(3.2,6.4,12.8,25.6μmol-L^-1)以及蝙蝠葛碱与3-MA联合用药对宫颈癌Hela细胞增殖的影响,应用流式细胞术观察自噬在蝙蝠葛碱诱导宫颈癌Hela细胞凋亡中的作用;Western blot检测蝙蝠葛碱和3-MA对宫颈癌Hela细胞Cleaved caspase-3、Bcl-2、Bax和Beclin-1表达的影响.结果蝙蝠葛碱显著抑制宫颈癌Hela细胞增殖,其抑制作用随着药物浓度的增加和作用时间的延长而增加;而3-MA与蝙蝠葛碱联合应用后,HeLa细胞凋亡率明显减少;蝙蝠葛碱上调Hela细胞Cleaved caspase-3、Bax和Beclin-1的表达(P<0.05),下调Bel-2蛋白表达(P<0.05);与蝙蝠葛碱组比较,3-MA与蝙蝠葛碱联合应用组Cleaved caspase-3、Beclin-1、Bax表达均明显降低(P<0.05),而Bel-2蛋白的表达明显升高(P<0.05):结论蝙蝠葛碱能够抑制宫颈癌HeLa细胞的增殖,上调凋亡相关蛋白Cleaved caspase-3、Bax和自噬相关蛋白Beclin-1的表达、下调抗凋亡蛋白Bcl-2表达,从而诱导宫颈癌HeLa细胞凋亡和自噬性死亡。
Objective To investigate the effect of autophagy in the apoptosis of cervical cancer Hela cells induced by Dauricine.Methods(1)MTT colorimetric assay detected the effects of Dauricine(3.2μM/L,6.4μM/L,12.8μM/L,25.6μM/L)and the combination of Dauricine and its autophagy inhibitor 3-MA in theproliferation of cervical cancer Hela cells.(2)The effect of autophagy in the apoptosis of cervical cancer Hela cells induced by Dauricine was observed by FLOW CYTOMETRY.(3)Western Blot detected the effects of Dauricine and its autophagy inhibitor 3-MA in the expression ol Cleaved caspase-3,Bel-2,Bax and Beclin-1 in cervical cancer Helacells.Results Dauricine significantly inhibited the proliferation of cervical cancer Hela cells,and its inhibitoiy effect increased with the ackl of drug concentration and prolonged action time;On the contraiy,the combination of 3-MA and Dauricine significantly decreased the apoptosis rate of He La cells;Dauricine up-regulated gene the expression of Cleaved caspase-3,Bax and Beclin-1 in Hela cells(P<0.05),anddown-regulated gene the expression of Bel-2(P<0.05).Compared with thegroup of Dauricine,3-MA combined withDaurieine the expressions of Cleaved caspase-3,Beclin-1 and Bax were significantly decreased(P<0.05).Bui the expression of Bel-2 was significantly increased(P<0.05).Conclusion Dauricine inhibits the proliferation of cervical cancer HeLa cells,lip-regulates gene the expression of apoptosis-related proteins Cleaved caspase-3,Bax and autophagy-related protein Beclin-1.And down-regukites gene the expression of anti-apoptotic protein Bel-2,thereby induced apoptosis and autophagic death of cervical cancer HeLci cells.
作者
元熙哲
窦浩文
陈世泽
李香丹
YUAN Xi-zhe;DOL Hao-wen;CHEN Shi-ze;LI Xiang-dan(Affiliated Hospital of Yanbian University,Yanji Jilin 133002,China;Medical College of Yanbian University,Yanji Jilin 133000,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2019年第12期3031-3033,共3页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81860461)。
