过表达CD44v6基因对人大肠癌SW480细胞侵袭迁移能力的影响  

Impact of CD44v6 overexpression on invasion and metastasis of colon cancer SW480 cells

作　　者：吕林[1] 刘海光[1] 张筱骅[1] 

机构地区：[1]温州医科大学附属第一医院肿瘤外科,浙江省温州市325000

出　　处：《世界华人消化杂志》2016年第8期1175-1183,共9页World Chinese Journal of Digestology

基　　金：温州市科技局基金资助项目;No.Y20140713;温州医科大学附属第一医院科研孵化基金资助项目;No.FHY2014013~~

摘　　要：目的:研究CD44v6基因过表达对SW480细胞侵袭和迁移能力的影响.方法:慢病毒介导的CD44v6过表达细胞(CD44v6组)和空载体对照细胞(NC组)由前期实验构建.采用荧光显微镜观察增强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)表达、实时荧光定量PCR检测CD44v6 mRNA表达水平、免疫荧光检测Flag标签蛋白三种方法重新鉴定过表达细胞模型;CCK-8法检测细胞增殖活性;划痕试验和Transwell试验检测细胞侵袭和迁移能力.结果:绿色荧光蛋白观察显示细胞转染效率近100%;实时荧光定量PCR显示CD44v6组细胞CD44v6 mRNA表达水平较对照组.显著升高(P<0.001);Flag标签蛋白免疫荧光染色显示过表达CD44v6蛋白主要定位于细胞膜.CCK-8结果显示2组细胞增殖无明显差异;划痕试验结果显示CD44v6组细胞划痕愈合指数较对照组显著增高(P<0.05);Transwell试验结果显示CD44v6组细胞迁移和侵袭相关指数均较对照组显著增高(均P<0.05),且CD44v6抗体处理后,CD44v6组细胞迁移和侵袭相关指数均较前显著减低(均P<0.05).结论:CD44v6基因过表达能显著增强SW480细胞侵袭和迁移能力.AIM： To investigate the impact of CD44v6 overexpression on the invasion and metastasis of human colon cancer SW480 cells.METHODS： SW480 cells stably overexpressing CD44v6 （CD44v6 group） and negative control cells （NC group） were developed through lentivirus infection. Transfection efficiency was evaluated by detecting the expression of enhanced green fluorescent protein （EGFP）. CD44v6 mRNA levels were determined using quantitative real-time PCR. Localization of the overexpressed protein was observed by immunofluorescence staining of Flag prot;ein, Cell proliferation was determined by cell counting kit （CCK）-8 assay. Cell invasion and metastasis were examined by scratch assay and transwell assay.RESULTS： EGFP detection indicated that transfection efficiency was close to 100% in both groups. CD44v6 mRNA levels in CD44v6 overexpressing cells were significantly higher than those in the control cells （P 〈 0.001）. The overexpressed CD44v6 protein was mainly localized on the cell membrane. No difference was revealed in cell proliferation between the two groups by CCK-8 assay. Scratch assay showed that the wound healing index was significantly increased in the CD44v6 group compared with the NC group （P 〈 0.05）.Transwell assay showed that the invasion and metastasis index was significantly increased in the CD44v6 group compared with the NC group （P 〈 0.05）, and the index was significantly decreased after the cells in the CD44v6 group were treated with CD44v6 antibody （P 〈 0.05）.CONCLUSION： Overexpression of CD44v6 significantly enhances the invasion and metastasis ability of SW480 cells.

关 键 词：大肠癌 侵袭 转移 CD44V6 过表达 

分 类 号：R735.34[医药卫生—肿瘤]