龙胆苦苷对人卵巢癌细胞株A2780/DDP顺铂化疗敏感性的影响及机制探讨  

Effect of Gentiopicroside on Chemosensitivity of Human Ovarian Cancer Cell Line A2780/DDP to Cisplatin and Its Mechanism

作　　者：井晶 刘梦 郭静[1] 田二斌 JING Jing;LIU Meng;GUO Jing;TIAN Erbin(The Second Affiliated Hospital of Zhengzhou University,Zhengzhou Henan China 450003)

机构地区：[1]郑州大学第二附属医院,河南郑州450003

出　　处：《中医学报》2021年第7期1506-1511,共6页Acta Chinese Medicine

基　　金：河南省教育厅科学技术研究重点项目(15A360 027)。

摘　　要：目的:探究龙胆苦苷对人卵巢癌细胞株A2780/DDP顺铂化疗敏感性的影响及其作用机制。方法:将耐顺铂人卵巢癌细胞株A2780/DDP进行细胞培养,分为4组,对照组、顺铂组、龙胆苦苷组、顺铂+龙胆苦苷组,顺铂组加入5 mg·L^(-1)顺铂,龙胆苦苷组加入15μmol·L^(-1)龙胆苦苷,顺铂+龙胆苦苷组加入15μmol·L^(-1)龙胆苦苷和5 mg·L^(-1)顺铂,对照组使用等体积的生理盐水。药物干预24 h后,光镜下观察A2780/DDP细胞形态;MTT法检测细胞增殖抑制率。药物干预48 h后,MTT法检测细胞增殖抑制率;TUNEL法检测细胞凋亡指数;RT-qPCR检测细胞中Bcl-2、p53、XIAP、Survivin的mRNA相对表达量;Western blot检测Bcl-2、p53、XIAP、Survivin蛋白表达水平及p53磷酸化水平。结果:与对照组相比,药物干预24 h后,A2780/DDP细胞形态出现改变,顺铂+龙胆苦苷组细胞形态改变最严重;A2780/DDP细胞增殖抑制率显著升高(P<0.05)。与对照组相比,药物干预48 h后,A2780/DDP细胞增殖抑制率显著升高,且高于24 h时的增殖抑制率(P<0.05);细胞凋亡指数显著升高(P<0.05);p53 mRNA水平显著升高(P<0.05);p53蛋白表达水平及磷酸化水平显著升高(P<0.05);Bcl-2、XIAP、Survivin的mRNA及蛋白表达水平显著降低(P<0.05)。结论:龙胆苦苷可抑制人卵巢癌细胞株A2780/DDP增殖,促进细胞凋亡,并可增强人卵巢癌细胞株A2780/DDP顺铂化疗敏感性,其机制可能与上调p53表达水平及磷酸化水平,下调Bcl-2、XIAP、Survivin表达水平有关。Objective:To investigate the effect of gentiopicroside on chemosensitivity of human ovarian cancer cell line A2780/DDP to cisplatin and its mechanism.Methods:Human ovarian cancer cell line A2780/DDP was cultured and divided into four groups:control group,cisplatin group,gentiopicroside group and cisplatin+gentiopicroside group.The cisplatin group was added with 5 mg·L^(-1) cisplatin,the gentiopicroside group was added with 15μmol·L^(-1) gentiopicroside,and the cisplatin+gentiopicroside group was added with 15μmol·L^(-1) gentiopicroside and 5 mg·L^(-1) cisplatin.The control group used the same volume of normal saline.After 24 hours of drug intervention,the morphology of A2780/DDP cells was observed under light microscope,and the inhibition rate of cell proliferation was detected by MTT method.After 48 hours of drug intervention,the inhibition rate of cell proliferation was detected by MTT method;Apoptosis index was detected by TUNEL;RT-qPCR was used to detect the relative mRNA expression of Bcl-2,p53,XIAP and Survivin;Western blot was used to detect the expression of Bcl-2,p53,XIAP,Survivin and the phosphorylation of p53.Results:Compared with the control group,the morphology of A2780/DDP cells changed after 24 hours of drug intervention,and the most serious change was in cisplatin+gentiopicroside group;the proliferation inhibition rate of A2780/DDP cells was significantly increased(P<0.05).Compared with the control group,the proliferation inhibition rate of A2780/DDP cells increased significantly after 48 hours of drug intervention,and was higher than that of 24 hours(P<0.05);the apoptosis index was significantly increased(P<0.05);the level of p53 mRNA was significantly increased(P<0.05);the expression level and phosphorylation level of p53 protein were significantly increased(P<0.05);The mRNA and protein expression levels of Bcl-2,XIAP and Survivin were significantly reduced(P<0.05).Conclusion:Gentiopicroside can inhibit the proliferation of human ovarian cancer cell line A2780/DDP,promote cell apoptosis,an

关 键 词：龙胆 龙胆苦苷 卵巢癌细胞 顺铂耐药 增殖抑制 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]