过表达连接蛋白32对人肝癌细胞株MHCC-97H体外侵袭能力的影响

Effect of Connexin 32 Overexpression on Invasion of Human Hepatocellular Carcinoma Cell Line MHCC-97H in vitro

目的探讨过表达连接蛋白32(Cx32)对人肝癌细胞株MHCC-97H体外侵袭能力的影响。方法应用Lipo-fectamine 2000将pcDNA 3.1(-)/Cx32重组质粒转染至MHCC-97H细胞,G418稳定筛选。采用Western blot法检测Cx32蛋白表达,划痕标记荧光传输实验显示细胞通讯功能的变化,Transwell实验检测细胞侵袭能力的改变。结果转染pcDNA 3.1(-)/Cx32重组质粒使得MHCC-97H细胞过表达Cx32蛋白。与正常对照组和转染空质粒组细胞相比,转染pcDNA 3.1(-)/Cx32重组质粒并过表达Cx32的细胞胞间通讯功能增强,细胞侵袭能力减弱。结论 Cx32在人原发性肝癌的侵袭过程中可能起抑制作用。

Objective To investigate the effect of Connexin 32（Cx32）on the invasion of human hepatocellular carcinoma cell line MHCC-97H. Methods The peDNA 3.1 （-）/Cx32 plasmid was transfeeted into MHCC-97H ceils. Cells with Cx32 overexpres sion were screened with G418 and validated by Western blot analysis, The gap junction intercellular communication between cells was detected by scrape-loading and dye transfer assay. The invasion of cells was detected by Transwell assay. Results The cells transfected with pcDNA 3.1 （-）/Cx32 plasmid overexpressed Cx32. As compared with blank controls and cells transfect ed with vector,the gap junction intercellular communication of cells with Cx32 overexpression was enhanced, while the number of migrating cells was decreased. Conclusion Cx32 may depress invasion of human hepatocellular carcinoma.