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目的探讨沉默血管内皮钙黏蛋白(vascular endothelial cadherin,VE-cadherin)基因对人高转移肝癌HCCLM3细胞增殖及迁移的影响。方法将携带3种不同抑制靶点的VE-cadherin基因沉默重组质粒GV248-shRNA-VEcadherin(1)、(2)、(3)分别转染至人高转移肝癌HCCLM3细胞,同时设阴性对照组(转染空载体质粒GV248-shRNANC)和空白对照组(未转染)。采用qRT-PCR及Western blot法分别检测各组细胞中VE-cadherin基因mRNA转录及蛋白的表达水平,同时筛选出最有效的靶点。MTT法测定各组细胞的体外增殖能力;Transwell小室试验检测各组细胞的迁移能力。结果与空白对照组及阴性对照组比较,3组转染VE-cadherin-shRNA干扰质粒的HCCLM3细胞的VE-cadherin基因mRNA转录及蛋白表达水平显著降低(P<0.05),其中GV248-shRNA-VE-cadherin(1)的VEcadherin基因抑制率最高(P<0.05),筛选为最佳靶点;最佳靶点干扰组HCCLM3细胞的增殖及迁移能力均明显降低(P<0.05)。结论沉默VE-cadherin基因可显著抑制人高转移肝癌HCCLM3细胞的增殖及迁移。
Objective To investigate the effects of silencing vascular endothelial cadherin (VE-cadherin) gene on the proliferation and migration of HCCLM3 cells with high metastatic potential. Methods The VE-cadherin gene silencing recombinant plasmids GV248-shRNA-VEcadherin (1), (2) and (3) were transfected into human HCCLM3 cells with three different inhibitory targets, Transfection of empty vector plasmid GV248-shRNANC) and blank control group (untransfected). The mRNA and protein expression of VE-cadherin in each group were detected by qRT-PCR and Western blot respectively, and the most effective target was screened out. MTT assay in vitro proliferation of cells; Transwell chamber test to detect the migration of cells in each group. Results Compared with the blank control group and the negative control group, the mRNA and protein expressions of VE-cadherin in HCCLM3 cells transfected with 3 groups of VE-cadherin-shRNA interference plasmids were significantly decreased (P <0.05), of which GV248-shRNA-VE The inhibition rate of VEcadherin gene of -cadherin (1) was the highest (P <0.05), and the screening was the best target. The proliferation and migration ability of HCCLM3 cells in the optimal target interference group were significantly decreased (P <0.05). Conclusion Silencing VE-cadherin gene can significantly inhibit the proliferation and migration of human HCCLM3 cells with high metastatic potential.