NEDD9表达下调对结肠癌生物学行为的影响

Effect of Down-regulating NEDD9 Expression on Biological Behavior of Colon Cancer

背景:NEDD9在细胞迁移、趋化、凋亡、细胞周期中发挥重要作用,NEDD9 siRNA可有效降低基因表达,并从不同水平上促进细胞凋亡、阻断肿瘤细胞迁移和侵袭。目的:探讨NEDD9在结肠癌组织中的表达以及siRNA干扰NEDD9表达对结肠癌Caco-2细胞增殖、凋亡、迁移和侵袭的影响。方法:收集2017年1月—2018年1月漯河市郾城区人民医院确诊的结肠癌组织和相应癌旁组织。常规培养结肠癌Caco-2细胞,采用Lipofectamine TM 2000转染细胞,并将细胞分为对照组、阴性对照组和siRNA干扰组。采用qRT-PCR法检测结肠癌组织和细胞中NEDD9 mRNA表达,CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,Transwell实验检测细胞迁移和侵袭,蛋白质印迹法检测NEDD9、MMP-9、Bcl-2、Bax、TIMP1蛋白表达。结果:结肠癌组织NEDD9 mRNA表达显著高于癌旁组织(P<0.05)。与对照组相比,siRNA干扰组NEDD9 mRNA表达明显降低(P<0.05),细胞增殖、迁移和侵袭能力受到明显抑制(P<0.05),细胞凋亡增强(P<0.05),NEDD9、MMP-9、Bcl-2蛋白表达显著降低(P<0.05),Bax、TIMP1蛋白表达显著增加(P<0.05)。结论:NEDD9基因可通过凋亡相关基因Bcl-2和Bax以及侵袭相关基因MMP-9和TIMP1来调控结肠癌Caco-2细胞增殖、凋亡、迁移和侵袭能力。

Background:NEDD9 plays an important role in cell migration,chemotaxis,apoptosis,and cell cycle.NEDD9 siRNA can effectively reduce the gene expression,promote apoptosis and block tumor cell migration and invasion at different levels.Aims:To investigate the expression of NEDD9 in colon cancer tissue,and the effect of NEDD9 siRNA on proliferation,apoptosis,migration and invasion of colon cancer Caco-2 cells.Methods:Colon cancer tissue and corresponding paracancerous tissue from Jan.2017 to Jan.2018 at Yancheng District People’s Hospital of Luohe were collected.Colon cancer Caco-2 cells were conventionally cultured,and were transfected by Lipofectamine TM 2000.Caco-2 cells were divided into control group,negative control group and siRNA interference group.The mRNA expression of NEDD9 in colon cancer tissue and Caco-2 cells was detected by qRT-PCR.The proliferation of Caco-2 cells was detected by CCK-8 method.The apoptosis of Caco-2 cells was detected by flow cytometry.Caco-2 cell migration and invasion were detected by Transwell assay.The protein expressions of NEDD9,MMP-9,Bcl-2,Bax,TIMP1 were determined by Western blotting.Results:The mRNA expression of NEDD9 in colon cancer tissue was significantly higher than that in paracancerous tissue(P<0.05).Compared with control group,the mRNA expression of NEDD9 in the siRNA interference group was significantly decreased(P<0.05),cell proliferation,migration and invasion were significantly inhibited(P<0.05),cell apoptosis was significantly increased(P<0.05),and the protein expressions of NEDD9,MMP-9 and Bcl-2 were significantly decreased(P<0.05),the protein expressions of Bax and TIMP1 were significantly increased(P<0.05).Conclusions:NEDD9 gene can regulate the proliferation,apoptosis,migration and invasion of colon cancer Caco-2 cells through apoptosis-related genes Bcl-2 and Bax and invasion-related genes MMP-9 and TIMP1.