吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (01): 83-89.doi: 10.13481/j.1671-587x.20180116

• 基础研究 • 上一篇    下一篇

五味子脂A增强卡铂对人卵巢癌Skov3细胞的促凋亡作用及其机制

上官梦原1, 赵菁2, 陈俊宇2, 杨艳荣3, 张佳悦2, 赵淑华1   

  1. 1. 吉林大学第二医院妇产科, 吉林 长春 130041;
    2. 延边大学附属医院妇产科, 吉林 延吉 133000;
    3. 吉林大学基础医学院病理生理学系, 吉林 长春 130021
  • 收稿日期:2017-08-31 出版日期:2018-01-28 发布日期:2018-01-24
  • 通讯作者: 赵淑华,教授,硕士研究生导师(Tel:0431-88796567,E-mail:zhaoshuhua-1966@163.com) E-mail:zhaoshuhua-1966@163.com
  • 作者简介:上官梦原(1987-),女,吉林省松原市人,医学硕士,主要从事妇科肿瘤治疗方面的研究。
  • 基金资助:
    吉林省科技厅科技发展计划项目资助课题(20140203013YY)

Enhancement of gomisin A in pro-apoptotic effect of carboplatin on human ovarian cancer Skov3 cells and its mechanism

SHANGGUAN Mengyuan1, ZHAO Jing2, CHEN Junyu2, YANG Yanrong3, ZHANG Jiayue2, ZHAO Shuhua1   

  1. 1. Department of Gynaecology and Obstetrics, Second Hospital, Jilin University, Changchun 130041, China;
    2. Department of Gynaecology and Obstetrics, Affiliated Hospital, Yanbian University, Yanji 133000, China;
    3. Department of Pathophysiology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China
  • Received:2017-08-31 Online:2018-01-28 Published:2018-01-24

摘要: 目的:探讨五味子脂A(GA)与卡铂(CBP)联合应用对卵巢癌Skov3细胞凋亡的影响,阐明GA的协同抑瘤作用。方法:采用不同浓度GA和CBP单独及联合对卵巢癌Skov3细胞株进行干预,MTT法检测各组细胞增殖抑制率,根据其结果选择GA和CBP合适浓度。卵巢癌Skov3细胞分为对照组、GA(0.04μmol·L-1)组、CBP(16mg·L-1)组、GA(0.04 μmol· L-1)联合CBP(16mg·L-1)组(联合用药组),药物处理48h后,观察各组细胞形态,流式细胞术检测细胞凋亡率,TUNEL染色观察细胞凋亡指数(AI),JC-1染色观察细胞线粒体膜电位变化,RT-PCR和Western blotting法检测凋亡相关基因Bax、ccaspase-3和Stat3 mRNA和蛋白表达水平。结果:GA联合CBP作用后细胞增殖抑制率高于GA和CBP单独应用组(P<0.01),且GA及CBP浓度分别为0.04 μmol·L-1和16 mg·L-1时量效比最佳(细胞增殖抑制率为52.1%)。与对照组比较,GA组和CBP组细胞折光性减弱,细胞回缩明显,部分脱落呈悬浮状;联合用药组细胞回缩现象更为明显,并见大量脱落细胞。流式细胞术和TUNEL检测,与对照组、GA组和CBP组比较,联合用药组Skov3细胞凋亡率和AI均明显升高(P<0.05或P<0.01)。JC-1染色检测,与GA组和CBP组比较,联合用药组Skov3细胞线粒体膜电位降低。RT-PCR和Western blotting法检测,联合用药组细胞中Bax和caspase-3mRNA和蛋白表达水平高于对照组、GA组和CBP组(P<0.05),Bcl-2mRNA和Stat3 mRNA及蛋白表达水平低于对照组、GA组和CBP组(P<0.05)。结论:GA能增强CBP对卵巢癌Skov3细胞的促凋亡作用,其机制可能与上调Bax和caspase-3的基因表达、下调Bcl-2的基因表达有关,可协同CBP诱导细胞凋亡。

关键词: 卵巢肿瘤, 五味子脂A, 卡铂, 细胞凋亡

Abstract: Objective: To explore the effect of gomisin A (GA) in combination with carboplatin (CBP)on the apoptosis of ovarian cancer Skov3 cells,and to illustrate the synergistic antitumor effect of GA. Methods: The ovarian cancer Skov3 cells were treated with GA combined with CBP.MTT assay was used to detectthe inhibitory rates of proliferation and the appropriate concentrations of GA and CBP were selected according to the results.The Skov3 cells were divided into control group,GA (0.04 μmol·L-1) group,CBP (16 mg·L-1)group and GA(0.04 μmol·L-1)+CBP(16 mg·L-1) group. After 48 h treatment,the cell morphology was observed by microscope,the apoptotic rate was measured by flow cytometry, the apoptotic index(AI) was observed by TUNEL staining,the mitochondrial membrane potential was detected by JC-1 staining,and the mRNA and protein expression levels of apoptosis-related genes (Bax,caspase-3,Stat3) were determined by RT-PCR and Western blotting method. Results: The inhibitory rate of proliferation of Skov3 cells in GA + CBP group(52.1%) was significantly higher than those in GA and CBP groups(P<0.01).The concentrations of GA and CBP for the best dose ratio were 0.04 μmol·L-1 and 16 mg·L-1,respectively. Compared with control group,the cell refractivities were decreased,the cells retracted,and part of cells were suspended in GA group and CBP groups;there were more suspended cells and cell retraction phenomenon was more prominent in GA + CBP group.The results of TUNEL staining and flow cytometry showed that the AI and apoptotic rate of cells in GA + CBP group were significantly increased compared with control group,GA and CBP group(P<0.05 or P<0.01);the JC-1 staining results suggested that the mitochondrial membrane potential of Skov3 cells in GA + CBP group was decreased(P<0.05 or P<0.01).The RT-PCR and Western blotting results showed that the expression levels of Bax and caspase-3 mRNA and protein were up-regulated(P<0.05) and the expression levels of Bcl-2 and Stat3 mRNA and protein in GA + CBP group were down-regulated compared with control,GA and CBP groups(P<0.05). Conclusion: GA can enhance the ability of CBP to induce the apoptosis of human ovarian cancer Skov3 cells,and its mechanisms may be related to the up-regulation of the Bax and Caspase-3 expressions and the down-reguation of the Bcl-2 expression;GA can synergize the induction of CBP on apoptosis.

Key words: gomisin A, apoptosis, ovarian neoplasms, carboplatin

中图分类号: 

  • R737.31