基因沉默PRDX1表达对人结直肠癌SW480细胞侵袭迁移的影响  

Effects of PRDX1 gene silencing on invasion and migration of human colorectal cancer SW480 cells

作　　者：冯继红[1] 张红[1,2] 李龙梅[1,2] 罗军敏[1,2] 臧春宝[1,3] 周航[1] 

机构地区：[1]遵义医学院附属医院肿瘤医院腹部肿瘤科,遵义563003 [2]遵义医学院免疫学教研室,贵州省免疫学研究生教育创新基地,遵义563003 [3]安徽省肿瘤医院放疗科,合肥230031

出　　处：《中国免疫学杂志》2017年第7期1048-1052,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81560407);贵州省科技厅社会发展攻关项目(黔科合SY字[2015]3046号);贵州省科技厅联合基金项目(黔科合LH字[2015]7505号);贵州省科学技术基金项目(黔科合J字[2013]2312号)资助

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〈O. O1 ）, 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 ceils 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.

关 键 词：过氧化还原酶1 结直肠癌 侵袭迁移 基质金属蛋白酶 SW480 

分 类 号：R735.3[医药卫生—肿瘤]