鱼藤素联合喜树碱对宫颈癌细胞自噬、凋亡及PI3K/Akt通路的影响  

Effects of Deguelin Combined with Camptothecin on Autophagy,Apoptosis and PI3K/Akt Pathway in Cervical Cancer Cells

作　　者：孔庆新 李思阳[2] 张径舟 罗丽梅 徐伟平 KONG Qingxin;LI Siyang;ZHANG Jingzhou;LUO Limei;XU Weiping(School of Pharmaceutical Engineering,Chongqing Vocational College of Chemical Technology,Chongqing 401228,China;School of Pharmacy,Jiangsu Food and Pharmaceutical Scinence College,Huai’an 223023 Jiangsu,China;Department of Laboratory,Southwest University of Political Science&Law Hospital,Chongqing 401120,China)

机构地区：[1]重庆化工职业学院制药工程学院,重庆401228 [2]江苏食品药品职业技术学院药学院,江苏淮安223023 [3]西南政法大学医院检验科,重庆401120

出　　处：《中药新药与临床药理》2022年第7期882-888,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：江苏省高校自然科学研究面上项目(17KJB360002)。

摘　　要：目的探讨鱼藤素联合喜树碱对宫颈癌HeLa细胞自噬、凋亡及PI3K/Akt通路的影响。方法以不同浓度鱼藤素(0、10、100、500、1000 nmol·L^(-1))、喜树碱(0、1、5、10、50μmol·L^(-1))及联合干预处理HeLa细胞48 h后,采用CCK-8法检测细胞增殖情况。将细胞分为对照组、鱼藤素组(100 nmol·L^(-1))、喜树碱组(5μmol·L^(-1))、联合干预组(100 nmol·L^(-1)鱼藤素+5μmol·L^(-1)喜树碱)、联合干预+LY294002组(100 nmol·L^(-1)鱼藤素+5μmol·L^(-1)喜树碱+5μg·mL^(-1)PI3K特异性抑制剂LY294002),分别进行相应干预48 h。采用流式细胞术检测HeLa细胞凋亡情况;采用透射电镜观察HeLa细胞自噬小体;Western Blot法检测HeLa细胞中凋亡、自噬及PI3K/Akt通路相关蛋白的表达。构建HeLa移植瘤裸鼠模型,分为对照组、鱼藤素组(10 mg·kg^(-1))、喜树碱组(6 mg·kg^(-1))、联合干预组(10 mg·kg^(-1)鱼藤素+6 mg·kg^(-1)喜树碱)及联合干预+LY294002组(10 mg·kg^(-1)鱼藤素+6 mg·kg^(-1)喜树碱+6 mg·kg^(-1)LY294002),连续干预21 d后检测肿瘤质量并计算抑瘤率。结果HeLa细胞增殖抑制率随鱼藤素和喜树碱浓度的增加而明显增加(P<0.05),联合干预组的细胞增殖抑制率明显高于鱼藤素、喜树碱单独处理组。与对照组相比,鱼藤素组、喜树碱组的HeLa细胞凋亡率明显升高(P<0.05),自噬小体数量明显增加(P<0.05),Caspase-3、Bax、Beclin1、LC3II/LC3I蛋白表达明显上调(P<0.05),p-PI3K/PI3K、p-Akt/Akt蛋白表达明显下调(P<0.05),HeLa移植瘤裸鼠肿瘤质量明显降低(P<0.05),抑瘤率明显升高。分别与鱼藤素组或喜树碱组相比,联合干预组的HeLa细胞凋亡率明显升高(P<0.05),自噬小体数量明显增加(P<0.05),Caspase-3、Bax、Beclin1、LC3II/LC3I蛋白表达明显上调(P<0.05),p-PI3K/PI3K、p-Akt/Akt蛋白表达明显下调(P<0.05),HeLa移植瘤裸鼠肿瘤质量明显降低(P<0.05),抑瘤率明显升高。与联合干预组相比,联合干预+LY294002组的HeLa细胞�Objective To investigate the effects of deguelin combined with camptothecin on autophagy,apoptosis and PI3K/Akt pathway in cervical cancer cells.Methods HeLa cells were treated with different concentrations of deguelin(0,10,100,500,1000 nmol·L^(-1))and camptothecin(0,1,5,10,50μmol·L^(-1))plus combined intervention for 48 hours,and CCK-8 method was adopted to detect the cell proliferation.The cells were divided into the control group,deguelin group(100 nmol·L^(-1)deguelin),camptothecin group(5μmol·L^(-1)camptothecin),combined intervention group(100 nmol·L^(-1)deguelin+5μmol·L^(-1)camptothecin),combined intervention+LY294002 group(100 nmol·L^(-1)deguelin+5μmol·L^(-1)camptothecin+5μg·mL^(-1)PI3K specific inhibitor LY294002),and the corresponding intervention was performed for 48 hours.The apoptosis of HeLa cells was detected by flow cytometry;the autophagosomes of HeLa cells were observed by transmission electron microscope;the protein expression of apoptosis,autophagy and PI3K/Akt pathway-related in HeLa cells were detected by Western Blot.The nude mouse model of HeLa transplanted tumor was constructed and divided into the control group,the deguelin group(10 mg·kg^(-1)),the camptothecin group(6 mg·kg^(-1)),the combined intervention group(10 mg·kg^(-1)deguelin+6 mg·kg^(-1)camptothecin)and the combined intervention+LY294002 group(10 mg·kg^(-1)deguelin+6 mg·kg^(-1)camptothecin+6 mg·kg-1 LY294002).The tumour mass was measured and tumour inhibition rate was calculated after 21 days of consecutive intervention.Results The proliferation inhibition rate of HeLa cells was significantly increased with the increase of deguelin and camptothecin concentrations(P<0.05),and the cell proliferation inhibition rate of the combined intervention group was significantly higher than that of the deguelin group and camptothecin group alone.Compared with the control group,the apoptosis rate of HeLa cells in the deguelin group and camptothecin group was significantly increased(P<0.05),the number of autophagic vesicles w

关 键 词：HELA细胞 鱼藤素 喜树碱 联合用药 凋亡 自噬 PI3K/AKT通路 

分 类 号：R285.5[医药卫生—中药学]