HOXB7基因沉默对人胃癌细胞活力、迁移和侵袭的影响

Effects of transcription factor HOXB7 silencing on viability,invasion and migration of SGC-7901 cells

目的:探讨靶向沉默HOXB7基因对人胃癌SGC-7901细胞活力、迁移和侵袭作用的影响及潜在机制。方法:设计靶向HOXB7的siRNA,然后瞬时转染胃癌SGC-7901细胞,实时荧光定量PCR和Western blot法检测siRNA靶向沉默的效果。MTT法检测细胞的活力,Western blot法检测细胞周期相关蛋白CDK4和cyclin D1蛋白的表达水平,Transwell小室侵袭实验检测SGC-7901细胞的侵袭能力,实时荧光定量PCR和Western blot法检测胃癌SGC-7901细胞PTEN和VEGF的表达水平以及p-AKT的蛋白水平。结果:胃癌组织中HOXB7基因在mRNA和蛋白水平的表达均显著高于对照组织。siRNA可有效靶向沉默SGC-7901细胞中HOXB7的表达,且沉默HOXB7表达后SGC-7901细胞活力明显下降,同时细胞周期相关蛋白CDK4和cyclin D1的表达亦下调。靶向沉默HOXB7可明显抑制SGC-7901细胞中AKT的磷酸化水平,抑制胃癌细胞的侵袭转移,同时下调VEGF的表达水平,抑制胃癌组织中的肿瘤血管生成。结论:HOXB7作为一个癌基因,可上调细胞周期相关蛋白CDK4、cyclin D1和侵袭相关分子VEGF的表达,上调AKT的磷酸化水平,进而抑制PTEN的功能,促进胃癌细胞SGC-7901的生长、迁移和侵袭能力。

AIM：To explore the effects of transcription factor HOXB7 silencing on the viability , invasion and migration of SGC-7901 cells.METHODS：The siRNA was designed to specifically interfere the expression of HOXB7.To assess the silence efficiency of the siRNA , the expression of HOXB7 at mRNA and protein levels was detected by real-time PCR and Western blot in the SGC-7901 cells transfected with siRNA .The cell viability was measured by MTT assay .Cell cycle-related proteins such as cyclin D 1 and CDK4 were determined by Western blot .Transwell assay was performed to analyze the migration ability of the SGC-7901 cells.The mRNA and protein levels of the tumor invasion-and metastasis-related molecules, such as VEGF, PTEN and p-AKT, were also detected by real-time PCR and Western blot .RESULTS：The expression of HOXB7 at mRNA and protein levels was higher in the gastric carcinoma tissues than that in the normal gastric tissues.The expression of HOXB7 at mRNA and protein levels was knockdown by siRNA .After silencing of HOXB7 gene expression, the viability of the SGC-7901 cells decreased, the expression of cyclin D1, CDK4 and VEGF was down-regulated, and the phosphorylation level of AKT was also obviously decreased .CONCLUSION：HOXB7 effectively promotes the viability, invasion and migration of gastric carcinoma cells by up-regulating the expression of cyclinD 1, CDK4 and VEGF, and increasing the phosphorylation level of AKT to inhibit PTEN function .