基因沉默PRDX1表达对人结直肠癌SW480细胞侵袭迁移的影响

摘要

目的:探讨RNA干扰过氧化还原酶1(Peroxiredoxin 1,PRDX1)表达对人结直肠癌SW480细胞侵袭转移能力的影响.方法:筛选RNA干扰PRDX1的慢病毒质粒,与阴性对照慢病毒质粒分组转染结直肠癌SW480细胞,转染后的SW480细胞可分为PRDX1基因沉默组(si-PRDX1)和阴性对照组(Vector).实时荧光定量PCR(qRT-PCR)和免疫印迹法(Western blot)分别检测两组细胞中 PRDX1 mRNA和蛋白表达;采用Transwell侵袭和迁移实验检测基因沉默PRDX1表达对结直肠癌细胞侵袭及迁移能力的影响;通过Western blot检测两组细胞中基质金属蛋白酶(MMP)家族部分蛋白表达水平.结果:基因沉默PRDX1表达可有效抑制结直肠癌SW480细胞中PRDX1 mRNA和蛋白水平的表达,与阴性对照组相比(Vector),差异均具有统计学意义(P<0.01),说明基因沉默PRDX1的SW480细胞系构建成功;Transwell侵袭和迁移实验显示si-PRDX1组细胞的侵袭及迁移能力较对照组均明显降低(P<0.01);Western blot结果显示,与Vector组相比,si-PRDX1组细胞中组织基质金属蛋白酶抑制剂2(TIMP-2)的表达明显增加,而MMP-2及MMP-9的表达显著下降,且差异均具有统计学意义(P<0.05).结论:基因沉默人结直肠癌SW480细胞的PRDX1表达可有效抑制细胞的侵袭、迁移及转移能力,其机制可能会通过调控TIMP-2、MMP-2及MMP-9的表达介导.%Objective:To investigate the effects of RNA interference(RNAi)-mediated silencing of Peroxiredoxin 1(PRDX1)gene on the invasion and migration of human colorectal cancer SW480 cells.Methods: Lentiviruses negative control vector and PRDX1 RNAi were transfected respectively into colorectal cancer SW480 cells.The transfected cells were divided into PRDX1 silencing group(si-PRDX1)and negative control group(Vector).The expressions of PRDX1 mRNA and protein in SW480 cells were exa mined by quantitative real-time PCR(qRT-PCR)and immunoblotting(Western blot),respectively.The cell migration and invasion capabilities were evaluated with transwell chamber assay and transwell chamber,respectively.The protein expressions of TIMP-2,MMP-2 and MMP-9 were detected by Western blot.Results: Compared with control group,the expressions of PRDX1 mRNA and protein were significantly decreased in PRDX1 silencing group(P<0.01),PRDX1 gene silencing cell line was successfully constructed.The levels of invasion and migration capacities of SW480 cells transfected with si-PRDX1 were lower than those in the cells transfected with control-siRNA(vector)(P<0.01).The expression of TIMP-2 was significantly increased,while the expressions of MMP-2 and MMP-9 were significantly decreased(P<0.05).Conclusion: Silencing of PRDX1 inhibits the invasion,migration and metastasis of human colorectal cancer SW480 cells by regulating the expressions of TIMP-2,MMP-2 and MMP-9.