PRDX2基因沉默促进结直肠癌细胞上皮-间质转化增强侵袭转移能力  

Silencing of PRDX2 promotes epithelial-mesenchymal transition of colorectal cancer cells and enhances the abilities of invasion and metastasis

作　　者：于伟娜[1] 冯继红[2] 丁陈波 高绍莹 徐林[1] 袁建波[3] 罗军敏[1] 

机构地区：[1]遵义医学院免疫学教研室暨贵州省免疫学研究生教育创新基地,贵州遵义563099 [2]遵义医学院附院附属肿瘤医院,贵州遵义563099 [3]遵义医学院医学检验系,2011级学生贵州遵义563099

出　　处：《遵义医学院学报》2015年第1期74-79,84,共7页Journal of Zunyi Medical University

基　　金：贵州省科学技术基金资助项目(NO:黔科合J字[2013]2312);贵州省优秀科技教育人才省长基金资助项目(NO:黔科教办[2010]04)

摘　　要：目的探讨Peroxiredoxin 2(PRDX2)基因沉默在结直肠癌细胞上皮-间质转化(EMT)中的作用及其参与侵袭转移的可能机制。方法将带有抗性的对照shRNA(shRNA-Control)和PRDX2 shRNA(shRNA-PRDX2)慢病毒颗粒分别转染结直肠癌SW480细胞,转染后细胞分为PRDX2沉默组(shRNA-PRDX2)和空载对照组(shRNA-Control)。采用免疫印迹法(Western blot)和实时荧光定量PCR(qRT-PCR)法分别检测两组细胞中PRDX2蛋白和mRNA表达;以5 ng/m L的TGF-β1作用两组细胞,在0、48、72 h观察两组细胞形态学变化;划痕实验和Transwell小室分别检测两组细胞体外迁移和侵袭能力;Western blot和qRT-PCR法检测E-cadherin、Vimentin、Snail、Twist1和MMP-9蛋白及mRNA表达。结果与对照组相比,PRDX2沉默组细胞PRDX2蛋白和mRNA的表达显著降低(P<0.05),成功建立PRDX2基因沉默稳定细胞株;经TGF-β1作用后,PRDX2沉默组细胞呈纺锤体样间质细胞表型,且迁移和侵袭能力明显增强;E-cadherin蛋白及mRNA水平表达显著下降,Vimentin、Snail、Twist1和MMP-9蛋白及mRNA表达水平均明显增加(P<0.05)。结论 PRDX2基因沉默可促进结直肠癌SW480细胞发生EMT,从而增强细胞的侵袭和转移能力,可能与其转录调节Snail、Twist1表达有关。Objective To explore the effects of PRDX2 silencing on epithelial - mesenchymal transition of color- ectal cancer cells, and its underlying mechanism in invasion and metastasis. Methods Lentiviruses control shR- NA （ shRNA - Control） and PRDX2 shRNA （ shRNA - PRDX2） with resistance were transfected respectively in- to coloreetal cancer SW480 cells. The transfected cells were divided into PRDX2 silencing group （shRNA - PRDX2） and empty vector control group （shRNA -Control）. The expressions of PRDX2 protein and mRNA in SW480 cells were examined by Immunoblot （ Western blot） and quantitative real - time PCR （ qRT - PCR）. Under the treatment with 5 ng/ml TGF - β1, cell morphological alterations were observed at 0, 48, and 72 h. The cell migration and invasion capabilities were evaluated with wound healing assay and transwell chamber, re- spectively. The protein and mRNA expressions of E - cadherin, Vimentin, Snail, Twistl and MMP - 9 were de- tected by Western blot and qRT - PCR. Results Compared with control group, the expressions of PRDX2 protein and mRNA were significantly decreased in PRDX2 silencing group （ P 〈 0.05 ）. PRDX2 silencing cell line was successfully constructed. The cells with PRDX2 silencing showed a spindle -like mesenchymal phenotype and the migration and invasion were significantly increased after treatment with TGF -β1. The expression of E - cad-herin was significantly decreased, while the expressions of Vimentin, Snail, Twistl and MMP -9 were signifi- cantly increased （ P 〈 0.05 ）. Conclusion Silencing of PRDX2 promotes epithelial - mesenchymal transition pos- sibly by regulating the expressions of Snail and Twistl in colorectal cancer SW480 cells, and subsequently pro- motes cell invasion and metastasis.

关 键 词：PEROXIREDOXIN 2 基因沉默 结直肠癌 上皮-间质转化 侵袭转移 

分 类 号：R735.3[医药卫生—肿瘤]