槲皮素通过内质网应激诱导宫颈癌细胞凋亡的机制  

Mechanism of quercetin inducing apoptosis of cervical cancer cell via endoplasmic reticulum stress

作　　者：脱淑梅[1] 李瑞瑞 李娟[1] 贾炎峰 李新茹[1] Tuo Shumei;Li Ruirui;Li Juan;Jia Yanfeng;Li Xinru(Gansu Provincial Maternity and Child-care Hospital,Lanzhou 730000,China)

机构地区：[1]甘肃省妇幼保健院,甘肃兰州730050

出　　处：《甘肃医药》2022年第9期769-773,共5页Gansu Medical Journal

基　　金：甘肃省卫生行业科研计划项目(编号:GSWSKY-2019-94)。

摘　　要：目的:分析槲皮素通过内质网应激对宫颈癌细胞凋亡的诱导情况,探讨其可能的作用机制。方法:将宫颈癌C33A细胞分为对照组和实验组,对照组(DMEM培养基+0.1%DMSO)、实验组(DMEM培养基+12.5、25.0、50.0、100μmol/L槲皮素)分别处理24h和48h。利用CCK8实验检测C33A细胞活力的变化,利用流式细胞技术检测细胞周期的变化,以实时定量PCR(qRT-PCR)技术检测内质网应激通路关键因子蛋白激酶R样内质网激酶(PERK)、真核翻译起始因子2α(eIF2α)、活化转录因子4(ATF4)、CCAAT增强子结合蛋白同源蛋白(CHOP)以及线粒体凋亡相关因子B淋巴细胞瘤-2基因(BAX)、半胱氨酸天冬氨酸特异性蛋白酶3(CASPASE3)、细胞色素C(CYT-C)和B细胞淋巴瘤/白血病-2(BCL-2)在转录水平的变化情况。结果:CCK8结果表明,经12.5、25.0、50.0、100μmol/L槲皮素处理24和48h后,C33A细胞活力均显著低于对照组,并且随着槲皮素浓度和时间的增加,细胞活力均呈下降趋势。流式细胞检测结果表明,槲皮素可增加C33A细胞周期G0/G1期数量,减少S和G2/M数量。qRT-PCR结果表明,槲皮素诱导内质网应激通路相关因子PERK、eIF2α、ATF4和CHOP的转录表达;同时细胞凋亡相关因子BAX、CASPASE3和CYT-C的转录表达也显著上调;与此相反,抗凋亡蛋白BCL-2的转录表达受到明显抑制。结论:槲皮素可通过内质网应激途径诱导宫颈癌细胞的细胞凋亡。Objective:To dissect the role of quercetin in inducing apoptosis of cervical cancer cell,and to understand the possible mechanism of this process.Methods:Cervical cancer cell C33A was divided into two groups,black group(DMEM medium+0.1%DMSO)and experimental group(DMEM medium+12.5,25.0,50.0,100μmol/L quercetin),and grown for 24 h and 48 h.CCK8 assay was used to detect the cell viability,flow cytometry was applied to test the cell cycle stage,quantitative real-time PCR(qRT-PCR)was employed to analyze the expression of endoplasmic reticulum stress-and apoptosis-related genes in C33A cells.Results:CCK8 results showed that the cell viability in experimental group was significantly reduced after treating with 12.5,25.0,50.0,100μmol/L quercetin for 24 and 48 h,comparing with that in black group.Moreover,the cell viability was gradually reduced via the time continued as well as the concentration of quercetin increased.flow cytometry results displayed that the number of G0/G1 stage cells was increased,the number of S and G2/M stage cells was decreased in experiment group,comparing with that in black group.qRT-PCR results showed that the expression of endoplasmic reticulum stress-related genes,PERK,eIF2α,ATF4 and CHOP,were all significantly induced by quercetin.Similarly,the expression of apoptosis-related genes,BAX,CASPASE3 and CYT-C were significantly induced by quercetin.In contrast,the expression of anti-apoptosis gene BCL-2 was significantly reduced by quercetin.Conclusion:Quercetin induces apoptosis of cervical cancer cell via endoplasmic reticulum stress.

关 键 词：槲皮素 宫颈癌细胞 内质网应激 细胞凋亡 

分 类 号：R285[医药卫生—中药学]