基础医学与临床 ›› 2015, Vol. 35 ›› Issue (12): 1596-1600.

• 研究论文 • 上一篇    下一篇

DADS通过miR-222抑制人胃癌细胞系的增殖与侵袭

黄宁江1,刘乐江2,盘箐3,张志伟4,唐海林5   

  1. 1. 永州职业技术学院药学系
    2. 永州职业技术学院
    3. 湖南永州职业技术学院医学院病原教研室
    4. 南华大学
    5. 南华大学肿瘤研究所
  • 收稿日期:2015-04-02 修回日期:2015-07-03 出版日期:2015-12-05 发布日期:2015-12-04
  • 通讯作者: 刘乐江 E-mail:176024235@qq.com
  • 基金资助:
    DADS上调miR-200b与miR-22表达抑制胃癌生长的分子机制研究

Diallyl Disulfide suppresses the proliferation and invasion of human gastric cancer cell line MGC-803 by down-regulating miR-222

  • Received:2015-04-02 Revised:2015-07-03 Online:2015-12-05 Published:2015-12-04

摘要: 目的 通过miRNA途径研究二烯丙基二硫(DADS)的抑瘤机制,以进一步阐明DADS抑制胃癌细胞增殖与转移的分子机制。方法 将胃癌细胞系MGC-803细胞分为DADS处理组、miR-222 mimics组、miR-222 inhibitors组和阴性对照组;分别采用0、 25、 50、100、 200和400μmol/L DADS处理MGC-803细胞;分别将miR-222 mimics、miR-222 inhibitors和scramble转染MGC-803细胞。qRT-PCR检测MGC-803细胞中miR-222表达;MTT法和Transwell侵袭实验检测MGC-803细胞增殖与侵袭能力;蛋白质印迹试验检测MGC-803细胞中TIMP3蛋白表达。结果 DADS可呈剂量依赖性下调MGC-803细胞中miR-222表达( P<0. 05);DADS能抑制MGC-803细胞的增殖与侵袭,外源高表达miR-222能促进MGC-803细胞的增殖与侵袭,而miR-222 inhibitors与DADS共同处理,MG-C803细胞的增殖与侵袭抑制作用最为显著( P<0. 05);DADS可下调MGC-803细胞中TIMP3蛋白的表达,外源高表达miR-222能上调MGC-803细胞中TIMP3蛋白的表达,而miR-222 inhibitors与DADS共同处理,MGC-803细胞中TIMP3蛋白的表达下调最为显著( P<0. 05)。结论 DADS通过下调miR-222的表达,靶向TIMP3抑制胃癌细胞的增殖与侵袭。

关键词: 胃癌, MiR-222, 二烯丙基二硫, 增殖与侵袭

Abstract: Objective: To identify the mechanism related to miRNA pathway which plays a role in the anti-tumor effects of Diallyl disulfide. Methods: Gastric cancer cell line MGC-803 were divided into DADS treatment group, miR-222 mimics group, miR-222 inhibitors group, negative control group; MGC-803 cells were treated With 0, 25, 50, 100, 200, 400μmol/L DADS respectively; miR-222 mimics, miR-222 inhibitors and scramble sequence were transfected in MGC-803 cells respectively. A qRT-PCR was employed for detecting the expression of miR-222 in MGC-803 cells. Western blotting was conducted to detect the expressions of TIMP3 protein in MGC-803 cells. The proliferation and invasion ability of MGC-803 cells in vitro were evaluated by MTT and Transwell invasion assays. Results: miR-222 expression was down- regulated with increasing doses of DADS( P<0. 05); DADS could significantly inhibit MGC-803 cell proliferation and invasion capacity, ectopic expression of miR-222 could significantly promote MGC-803 cell proliferation and invasion capacity. Furthermore, gastric cancer cells treated with combination of DADS and miR-222 inhibitors showed significant inhibition of cell proliferation and invasion than treated with DADS alone( P<0. 05); The level of TIMP3 protein was inhibited by treated with DADS in MGC-803 cells, overexpression of miR-222 resulted in increased TIMP3 protein levels. Furthermore, The level of TIMP3 protein was inhibited treated with combination of DADS and miR-222 inhibitors than treated with DADS when alone in MGC-803 cells( P<0. 05). Conclusion: DADS suppresses proliferation and invasion in human gastric cancer cells through targeting TIMP3 by down-regulation of miR-222.

Key words: Gastric cancer, MiR-222, Diallyl Disulfide, Proliferation and Invasion

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