吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (02): 292-298.doi: 10.13481/j.1671-587x.20180216

• 基础研究 • 上一篇    下一篇

五味子脂A联合卡铂对人卵巢癌Skov3细胞增殖、迁移和侵袭的抑制作用及其机制

上官梦原1, 赵菁2, 杨艳荣3, 张佳悦2, 陈俊宇1, 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:2018-03-28 发布日期:2018-03-30
  • 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)

Inhibitory effects of gomisin A combined with carboplatin on proliferation,invasion and metastasis of human ovarian cancer Skov3 cells and their mechanisms

SHANGGUAN Mengyuan1, ZHAO Jing2, YANG Yanrong3, ZHANG Jiayue2, CHEN Junyu1, 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:2018-03-28 Published:2018-03-30

摘要: 目的:探讨五味子脂A (GA)与卡铂(CBP)联用对人卵巢癌Skov3细胞增殖、迁移与侵袭的抑制作用及其机制,阐明GA的协同抑瘤作用。方法:采用不同浓度GA与CBP单独或联合对卵巢癌Skov3细胞株进行干预,MTT法检测细胞增殖抑制率,根据其结果选择GA和CBP合适浓度。实验分为对照组、GA (0.04 μmol· L-1)组、CBP (16mg· L-1)组、GA (0.04 μmol·L-1)联合CBP (16mg·L-1)组(GA+CBP组),药物处理48h后,克隆形成实验检测细胞克隆形成能力,细胞划痕实验检测各组细胞迁移能力,Transwell实验检测各组细胞侵袭能力,RT-PCR法检测各组细胞中基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9) mRNA表达水平,Western blotting法检测各组细胞中MMP-2、MMP-9、AKT1和pAKT1蛋白表达水平。结果:与GA组和CBP组比较,GA+CBP组细胞增殖抑制率明显升高(P<0.01),且GA和CBP浓度分别为0.04 μmol·L-1、16 mg·L-1时量效比最佳。克隆形成实验,与对照组比较,CBP组和GA+CBP组Skov3细胞克隆形成率均降低(P<0.05或P<0.01);GA+CBP组Skov3细胞形成率明显低于GA组和CBP组(P<0.01)。细胞划痕实验,与对照组比较,GA组和CBP组Skov3细胞迁移的数量减少;GA+CBP组细胞迁移的数量明显低于GA组和CBP组。Transwell实验,与对照组比较,GA组和CBP组Skov3细胞穿过细胞外基质(ECM)的能力降低;与GA组和CBP组比较,GA+CBP组Skov3细胞穿过ECM的能力降低。RT-PCR法,与GA组和CBP组比较,GA+CBP组Skov3细胞中MMP-2和MMP-9 mRNA表达水平明显降低(P<0.01);Western blotting法,与GA组和CBP组比较,GA+CBP组Skov3细胞中MMP-2、MMP-9、AKT1和pAKT1蛋白表达水平明显降低(P<0.01)。结论:GA可增强CBP抑制人卵巢癌Skov3细胞增殖、侵袭和转移的能力,从而发挥化疗的减毒增效作用。

关键词: 细胞增殖, 侵袭转移, 五味子脂A, 卡铂, 卵巢肿瘤

Abstract: Objective:To explore the inhibitory effects of gomisin A (GA) in combination with carboplatin (CBP)on the proliferation,invasion and metastasis of human ovarian cancer Skov3 cells and their mechanisms,and to illustrate the synergistic antitumor effect of GA. Methods: The ovarian cancer Skov3 cells were treated with different concentrations of GA combined with different concentrations of CBP.MTT assay was used to detect the inhibitory rates of proliferation.According to the results,the appropriate concentrations of GA and CBP were conformed.The Skov3 cells were treated with PBS,GA(0.04 μmol·L-1),CBP (16 mg·L-1) and GA (0.04 μmol·L-1) in combination with CBP (16 mg·L-1),respectively, and used as control group,GA group,CBP group and GA+CBP group. After 48 h of treatment,the cell morphology was observed by optical microscope,the cell reproductive ability was determined by clonogenic cell survival assay,and the cell migration ability was measured by cell wound healing test;Transwell experiment was used to detect the cell invasion ability.The expression levels of MMP-2 and MMP-9 mRNA were determined by RT-PCR and the protein expression levels of MMP-2,MMP-9,AKT1 and pAKT1 were determined by Western blotting method. Results: Compared with GA and CBP groups,the inhibitory rate of proliferation of Skov3 cells in GA + CBP group was significantly increased(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. The results of clone formation assay showed that the colony formation rates in CBP and GA+CBP groups were decreased compared with control group(P<0.05 or P<0.01);the colony formation rate of cells in GA + CBP group was significantly decreased compared with GA and CBP groups(P<0.01); the wound scratch assay results suggested that the number of metastic Skov3 cells in GA and CBP groups were decreased compared with control group;and the number of metastic Skov3 cells in GA + CBP group was decreased compared with GA and CBP groups.The results of Transwell expriment showed that the capacities of cells passing the ECM in GA and CBP groups were decreased compared with control group;the capacity of cells passing the ECM was decreased compared with GA and CBP groups.The RT-PCR and Western blotting results showed that the expression levels of MMP-2 and MMP-9 mRNA in GA+CBP group were down-regulated compared with GA and CBP groups(P<0.01),and the expression levels of MMP-2,MMP-9,AKT1 and pAKT1 protein were down-regulated(P<0.01). Conclusion: GA can enhance the ability of CBP to inhibit the proliferation,invasion and metastasis of human ovarian cancer Skov3 cells,and thus play a role in decreasing the toxicity and increasing the efficacy of chemotherapy.

Key words: carboplatin, invasion and metastasis, ovarian neoplasms, gomisin A, cell proliferation

中图分类号: 

  • R285.5