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

doi:10.13481/j.1671-587x.20180216

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

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. 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. 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/Abstract

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

上官梦原, 赵菁, 杨艳荣, 张佳悦, 陈俊宇, 赵淑华. 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[J]. 吉林大学学报(医学版), 2018, 44(02): 292-298.

SHANGGUAN Mengyuan, ZHAO Jing, YANG Yanrong, ZHANG Jiayue, CHEN Junyu, ZHAO Shuhua. Inhibitory effects of gomisin A combined with carboplatin on proliferation,invasion and metastasis of human ovarian cancer Skov3 cells and their mechanisms[J]. Journal of Jilin University Medicine Edition, 2018, 44(02): 292-298.

参考文献

[1] Waiwut P,Shin MS,Yokoyama S,et al.Gomisin A enhances tumor necrosis factor-α-induced G1 cell cycle arrest via signal transducer and activator of transcription 1-mediated phosphorylation of retinoblastoma protein[J].Biol Pharm Bull,2012,35(11):1997-2003.
[2] 孙雨薇,闫冬梅.五味子多糖对体外培养肝癌SMMC-7721细胞Bcl-2和Bax蛋白表达的影响[J].中国处方药,2016,14(7):21-22.
[3] 陈海芳,戚进,余伯阳.生脉散中人参、麦冬和五味子提取物体外抗氧化作用[J].中国实验方剂学杂志,2016,22(19):37-42.
[4] Leong PK,Chiu PY,Chen N,et al.Schisandrin B elicits a glutathione antioxidant response and protects against apoptosis via the redox-sensitive ERK/Nrf2 pathway in AML12 hepatocytes[J].Free Radical Res,2011,45(4):483-495.
[5] 安琳,苏慎勇,杨卫东,等.参芪五味子片口服降低胃癌化疗后胃肠道抑制及对患者免疫功能的影响[J].中药药理与临床,2017(2):226-229.
[6] Zhao T,Mao G,Mao R,et al.Antitumor and immunomodulatory activity of a water-soluble low molecular weight polysaccharide from Schisandra chinensis (Turcz.) Baill[J].Food Chem Toxicol,2013,55:609-616.
[7] Kim SJ,Min HY, Lee EJ,et al.Growth inhibition and cell cycle arrest in the G0/G1 by schizandrin,a dibenzocyclooctadiene lignan isolated from Schisandra chinensis,on T47D human breast cancer cells[J].Phytother Res,2010,24(2):193-197.
[8] 赖一鸣,许娜,陈晓杰,等.五味子多糖抑制卵巢癌SKOV3细胞增殖的研究[J].中华中医药学刊,2014(4):856-857.
[9] 上官梦原.五味子脂A增强紫杉醇和卡铂对Skov3的抑瘤作用及机制研究[D].长春:吉林大学,2014.
[10] 齐乃松,郭建,王雪,等.软琼脂克隆形成实验评价药物体外抑瘤性与成瘤性[J].药物分析杂志,2017,37(3):444-450.
[11] 章尤权,王清泰,陈旭征,等.白花蛇舌草对人肝癌HepG₂细胞裸鼠皮下移植瘤PI3K/Akt信号通路的影响[J].肿瘤基础与临床,2015,28(4):277-280.
[12] 刘文涓,王前,李凤杰,等.p-Akt-mTOR-p70S6K信号通路蛋白与卵巢癌临床病理特征及化疗耐药的相关性[J].中南大学学报:医学版,2017,42(8):882-888.
[13] 刘慧,李鑫,胡腾鹏,等.PI3K/AKT/mTOR信号通路抑制剂在淋巴瘤中的研究进展[J].中国肿瘤临床,2016,43(5):211-215.
[14] 曾莉,唐小珂,周琦.PI3K/AKT信号通路及抑制剂的研究进展[J].现代妇女:医学前沿,2015(1):309.
[15] Markman B,Dienstmann R,Tabernero J.Targeting the PI3K/Akt/mTOR pathway-beyond rapalogs[J].Oncotarget,2010,1(7):530-543.
[16] 张瑶.PI3K/AKT信号通路与人上皮性卵巢癌耐药性的相关性研究[D].南昌:南昌大学,2014.
[17] 喻学桥.细胞外基质重构对肿瘤浸润转移的影响[J].肿瘤学杂志,2015,21(5):428-431.
[18] Huo X,Liu C,Bai X,et al.Aqueous extract of Cordyceps sinensis potentiates the antitumor effect of DDP and attenuates therapy-associated toxicity in non-small cell lung cancer via IκBα/NFκB and AKT/MMP2/MMP9 pathways[J].RSC Advances,2017,7(60):37743-37754.
[19] 杨苏钰,唐德才,曹子丰,等.黄芪甲苷配伍姜黄素对人卵巢癌HO-8910原位移植瘤转移的抑瘤作用[J].中国实验方剂学杂志,2017,23(6):155-160.
[20] 张展,朱海,宋华,等.Lgr5、β-catenin在卵巢上皮性癌组织中的表达及对SKOV3细胞增殖迁移的影响[J].郑州大学学报:医学版,2017,52(2):138-142.
[21] 杨蓉,高婷婷,姚念玲,等.沉默IFITM1基因对卵巢癌CP70细胞增殖和侵袭能力的影响[J].西安交通大学学报:医学版,2016,37(4):544-546,577.

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

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

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