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RORα高表达对二烯丙基二硫抑制人胃癌细胞增殖与迁移侵袭的影响

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RORα高表达对二烯丙基二硫抑制人胃癌细胞增殖与迁移侵袭的影响
Effect of RORα overexpression on diallyl disulfide-induced suppression of proliferation, migration and invasion of human gastric MGC803 cells
上传时间:2019/8/23 17:54:17    作者:苏坚,夏红,曾颖,苏波,刘芳,凌晖,曾希,苏琦|下载

苏坚1. 2#,夏红1#,曾颖1,苏波1,刘芳1,凌晖1,曾希1,苏琦1*
(1南华大学附属第二医院病理科,衡阳 421001;2南华大学肿瘤研究所,湖南省肿瘤细胞与分子病理学重点实验室,湖南省胃癌研究中心,衡阳 421001)

Su Jian1,2#, Xia Hong 1#, Zhen ying1, Liu Fang 1, Su Bo 1, Ling Hui 1, Zhen Xi 1, Su Qi 1*
(1 Department of pathology, the second Affiliated Hospital, University of South China, Hengyang 421001, China; 2 Cancer Research Institute, University of South China, Hunan Provincial Key Laboratory of Cancer Cellular And Molecular Pathology, Center for Gastric Cancer Research of Hunan Province, Hengyang 421001, China)
摘要:目的 观察高表达RORα对二烯丙基二硫(DADS)抑制人胃癌MGC803细胞增殖、迁移与侵袭的影响。方法 集落形成实验与流式细胞术检测细胞增殖与细胞周期;细胞划痕和Transwell实验分别检测细胞迁移与侵袭。RT-PCR与Western blot分别检测RORα、MMP-9和TIMP3 mRNA与蛋白表达水平。结果RT-PCR与Western blot检测显示,RORα高表达与DADS处理较对照组与空载体组RORα mRNA与蛋白表达明显上调,DADS+RORα高表达组上调更为显著(P<0.05)。与对照组和空载体组比较,RORα高表达与DADS处理组MMP-9表达下调,TIMP3表达上调,DADS+RORα高表达组改变最为显著。集落形成实验显示,RORα高表达与DADS处理组较对照组与空载体组的集落形成率明显降低。流式细胞术显示,与对照组和空载体组比较,RORα高表达与DADS处理组G2/M期细胞比率明显升高。细胞划痕和Transwell实验显示,RORα高表达与DADS处理组细胞迁移与侵袭能力明显降低。结论RORα高表达可通过上调TIMP3与下调MMP-9促进DADS阻滞MGC803细胞G2/M期和抑制增殖与迁移侵袭。
关键词:RORα;二烯丙基二硫;人胃癌MGC803细胞;增殖;迁移;侵袭
Abstract:Objective   To study the effect of RORα overexpression on the proliferation, migration and invasion in human gastric cancer MGC803 cells suppressed by diallyl disulfide (DADS). Methods The colony formation and flow cytometry were used to detect cell proliferation and cell cycle. Scratch and Transwell assay were performed to examine cell migration and invasion. RORα, MMP-9 and TIMP3 mRNA and protein levels were detected by RT-PCR and Western blot, respectively. Results The expression of RORα in RORα overexpression and DADS treated group (DADS+RORα) was significantly up-regulated compared with that in the control group and the empty vector group, but the expression of MMP-9 was down-regulated in the DADS+RORα group, and the expression of TIMP3 was up-regulated, among which the change in the DADS+RORα group was the most significant.After DADS treated, the colony formation rate of each group was reduced, and RORα overexpression was significantly lower than that before treatment. The colony forming efficiency in DADS+RORα group was 34.90% lower than that in the control group and the empty vector group. Flow cytometry showed that the ratio of G2/M phase cells with RORα overexpression and DADS treatment was overtly higher than that of the control group and the empty vector group. Scratch and Transwell assay showed that RORα overexpression and DADS treatment significantly inhibited the migration and invasion of MGC803 cells. Conclusion RORα overexpression could promote the arrest of G2/M phase cells and inhibition of proliferation, migration and invasion by DADS through upregulating TIMP3 and downregulating MMP-9 in MGC803 cells .
Keywords: RORα; diallyl disulfide; human gastric cancer cells; proliferation; migration; invasion
〔中图分类号〕R735.2             〔文献标识码〕A             DOI:10.16705/ j. cnki. 1004-1850. 2019. 03. 006

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