槲皮素对宫颈癌Hela/DDP细胞顺铂耐药性的影响及作用机制研究  

Effect of quercetin on cisplatin resistance on cervical cancer HeLa/DDP cells and the underlying mechanism

作　　者：余琴 刘艳[1] 柯娟[1] 赵永艳 YU Qin;LIU Yan;KE Juan;ZHAO Yongyan(Department of Traditional Chinese Medicine,Taihe Hospital,Shiyan,Hubei 442000)

机构地区：[1]湖北省十堰市太和医院中医部,湖北十堰442000

出　　处：《河北中医》2025年第3期456-460,467,共6页Hebei Journal of Traditional Chinese Medicine

基　　金：十堰市太和医院科研项目(编号:2020JJXM105)。

摘　　要：目的探究槲皮素对宫颈癌Hela/DDP细胞顺铂耐药性的影响及其可能作用机制。方法体外培养人宫颈癌Hela/DDP、Hela细胞,按如下分组开展实验:(1)Hela/DDP、Hela细胞设置不同浓度(0、5、10、20、40、80μg/mL)顺铂处理组;(2)Hela/DDP细胞设置对照组(不处理)、顺铂组(10μg/mL顺铂)、低浓度槲皮素组(10μg/mL顺铂+20μmol/L槲皮素)、中浓度槲皮素组(10μg/mL顺铂+40μmol/L槲皮素)和高浓度槲皮素组(10μg/mL顺铂+80μmol/L槲皮素);(3)Hela/DDP细胞设置NC组、NC-siRNA组和miRNA-18a-siRNA组,按组转染处理,均加入终浓度为10μg/mL的顺铂和80μmol/L的槲皮素。CCK-8法检测Hela/DDP、Hela细胞增殖情况;流式细胞仪检测Hela/DDP细胞凋亡情况;实时荧光定量聚合酶链式反应法检测Hela/DDP细胞中miRNA-18a表达情况;蛋白免疫印迹法检测Hela/DDP细胞中B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达情况。结果使用5、10、20、40、80μg/mL浓度的顺铂处理后,Hela/DDP细胞增殖抑制率均低于Hela细胞(P<0.05);顺铂对Hela、Hela/DDP细胞的半数抑制浓度(IC50)分别处于5~10μg/mL、10~20μg/mL之间。与对照组相比,顺铂组Hela/DDP细胞增殖率、细胞中Bcl-2蛋白表达水平显著降低(P<0.05),凋亡率、细胞中miRNA-18a、Bax蛋白表达水平显著升高(P<0.05);与顺铂组相比,低、中、高浓度槲皮素组Hela/DDP细胞增殖率、细胞中Bcl-2蛋白表达水平降低(P<0.05),凋亡率、细胞中miRNA-18a、Bax蛋白表达水平升高(P<0.05),且呈剂量依赖性(P<0.05)。与NC-siRNA组相比,miRNA-18a-siRNA组Hela/DDP细胞中miRNA-18a表达水平及细胞凋亡率均显著降低(P<0.05),细胞光密度(OD)值显著升高(P<0.05)。结论槲皮素可在顺铂处理宫颈癌Hela/DDP细胞基础上进一步抑制细胞增殖并诱导细胞凋亡,且可能是通过上调miRNA-18a表达逆转宫颈癌细胞的顺铂耐药性。Objective To investigate the effect of quercetin on cisplatin resistance on cervical cancer cells Hela/DDP and its possible mechanism.Methods Hela/DDP and Hela cells were cultured in vitro,and perform experiments in groups as follows.First,Hela/DDP and Hela cells were treated with cisplatin at various concentrations(0,5,10,20,40,80μg/mL).Second,Hela/DDP cells were divided into control group(no treatment),cisplatin group(10μg/mL cisplatin),low-dose quercetin group(10μg/mL cisplatin+20μmol/L quercetin),medium-dose quercetin group(10μg/mL cisplatin+40μmol/L quercetin)and high-dose quercetin group(10μg/mL cisplatin+80μmol/L quercetin).Third,Hela/DDP cells were divided into NC group(no transfection),NC siRNA group(transfected with NC siRNA)and miRNA-18a-siRNA group(transfected with miRNA-18a-siRNA),followed by treatment with cisplatin(10μg/mL)and quercetin(80μmol/L).The proliferation of Hela/DDP and Hela cells was detected by CCK-8.assay.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression of miRNA-18a in Hela/DDP cells.The apoptosis of Hela/DDP cells was detected by flow cytometry.Western blot was used to detect the expressions of Bcl-2 and Bax in Hela/DDP cells.Results After cisplatin induction,the proliferation inhibition rate of Hela/DDP cells was significantly lower than that of Hela cells(P<0.05).The half inhibitory concentration(IC50)of cisplatin was 5-10μg/mL in Hela cells and 10-20μg/mL in Hela/DDP cells.Compared with those in the control group,the proliferation rate and protein expression of Bcl-2 in Hela/DDP cells of the cisplatin group were significantly lower(P<0.05),while the apoptosis rate and the expression levels of miRNA-18a and Bax were significantly higher(P<0.05).Quercetin treatment dose-dependently inhibited the proliferation rate and protein expression of Bcl-2 in Hela/DDP cells(P<0.05),but increased the apoptosis rate,and expression levels of miRNA-18a and Bax(P<0.05),and showing a dose-dependent(P<0.05).Compared with those in NC siRNA group,expression l

关 键 词：宫颈肿瘤 槲皮素 顺铂 抗药性 肿瘤 微RNAS BCL-2相关X蛋白质 药理作用分子作用机制(中药) 

分 类 号：R737.33[医药卫生—肿瘤] R730.52[医药卫生—临床医学] R73-37R286.91