沉默Ephrin B2基因表达对结直肠癌细胞侵袭和迁移的影响及其机制

Effects of silencing Ephrin B2 gene on invasion and migration of human colorectal carcinoma cells

目的探讨沉默肝配蛋白B2(Ephrin B2)基因表达对结直肠癌SW480细胞(以下称SW480细胞)侵袭和迁移的影响及其机制。方法体外培养SW480细胞,随机分为Ephrin B2组、阴性对照组、空白对照组。Ephrin B2组、阴性对照组分别转染Ephrin B2-siRNA、NC-siRNA,空白对照组不予转染。转染8 h再培养48 h,收集细胞。分别采用Transwell侵袭实验、划痕修复实验检测细胞侵袭和迁移能力,采用Western blotting法和qRT-PCR法检测E-cadherin、N-cadherin、Vimentin蛋白和mRNA表达。结果 Ephrin B2组细胞侵袭和迁移能力均明显低于阴性对照组和空白对照组(P均<0.05),而阴性对照组与空白对照组比较P均>0.05。与阴性对照组和空白对照组比较,Ephrin B2组N-cadherin、Vimentin蛋白和mRNA表达均明显降低,E-cadherin蛋白和mRNA表达均明显升高(P均<0.05),而阴性对照组与空白对照组比较P均>0.05。结论沉默Ephrin B2基因表达能抑制结直肠癌细胞侵袭和迁移能力,其机制可能与调控上皮间质转化过程有关。

Objective To investigate the effects of silencing Ephrin B2 gene expression on the invasion and migration of human colorectal carcinoma SW480 cells and to explore its possible mechanism. Methods SW480 cells were randomly divided into the Ephrin B2 group,negative control group（ NC group）,and blank control group（ CON group）. We transfected the Ephrin B2 RNAi and NC-siRNA into the Ephrin B2 group and negative control group by Lipofectamine2000 liposome; the bland control group was not transfected. We replaced the new medium after transfection for 8 h,continued to culture for 48 h and then collected the cells. The invasion and migration of SW480 cells were detected by Transwell assay and Scratch test.The expression levels of Ephrin B2 and EMT markers proteins（ E-cadherin,N-cadherin,and Vimentin） mRNA and protein were detected by Western blotting and real-time fluorescence quantitative PCR（ qRT-PCR）. Results The invasion and migration abilities of SW480 cells in the Ephrin B2 group were lower than those of the NC group and CON group,and the difference was statistically significant（ both P〈0. 05）. Compared with the NC group and CON group,the protein and mRNA expression levels of N-cadherin and Vimentin decreased significantly,and the protein and mRNA expression levels of E-cadherin increased in the Ephrin B2 group（ all P〈0. 05）,but no significant difference was found between the the NC group and CON group（ all P〈0. 05）. Conclusion Silencing Ephrin B2 expression can effectively inhibit the invasion and migration colorectal carcinoma cells,and its mechanism may be related to the regulation of the EMT process.