抑制KIAA1429表达对人胃癌细胞侵袭和迁移的影响及其机制

Effect of KIAA1429 knockdown on invasion and migration of human gastric cancer cells and its mechanism

目的观察抑制KIAA1429基因对胃癌细胞侵袭和迁移能力的影响,并探讨其机制。方法将细胞随机分为两组:干扰组、对照组,分别转染针对KIAA1429基因的干扰慢病毒及阴性对照空载慢病毒。分别采用qRT-PCR法和Western blot法检测细胞中的KIAA1429 mRNA 和蛋白表达情况;采用Transwell 实验观察细胞体外迁移及侵袭能力(穿膜细胞数),细胞划痕实验观察细胞体外迁移能力(24 h迁移距离);采用Western blot法检测细胞上皮间充质转化(EMT)相关分子(E-Cadherin、N-Cadherin)蛋白表达量。结果抑制KIAA1429基因表达后,KIAA1429 mRNA及蛋白表达量降低;与对照组比较,干扰组穿膜细胞数减少,24 h迁移距离减少,细胞中N-Cadherin蛋白表达减少,E-Cadherin蛋白表达增加( P <0.05)。结论慢病毒介导的抑制KIAA1429基因表达,可降低胃癌SGC7901和MGC803细胞迁移和侵袭能力;其机制可能与调控N-Cadherin、E-Cadherin的表达从而抑制EMT过程有关。

Objective To observe the effect of inhibiting KIAA1429 gene on the invasion and migration of gastric cancer cells, and to explore its mechanism. Methods The cells were randomly divided into two groups. The knockdown group and the control group were transfected with the interfering lentivirus against the KIAA1429 gene and the negative control empty lentivirus. The expression of KIAA1429 mRNA and protein was detected by quantitative real-time polymerase chain reaction(qRT-PCR) and Western blot. The migration ability was observed by cell scratch assay and Transwell assay. The invasion ability was observed by Transwell assay. Western blot was used to detect the expression of epithelial-mesenchymal transition(EMT)-related molecules(E-Cadherin, N-Cadherin). Results The expression of KIAA1429 mRNA and protein was decreased after inhibiting the expression of KIAA1429 gene. Compared with the control group, the number of transmembrane cells in the knockdown group was decreased, the migration distance was decreased at 24 h, the expression of N-Cadherin protein was decreased, the expression of E-Cadherin protein was increased( P <0.05). Conclusion Lentivirus-mediated inhibition of KIAA1429 gene expression can reduce the migration and invasion of gastric cancer SGC7901 and MGC803 cells. The mechanism may be related to the regulation of N-Cadherin and E-Cadherin expression and inhibition of EMT.