DDR1通过NF-KB调控胃癌细胞转移的机制研究

DDR1 promotes the migration and invasion of gastric cancer cell by up-regulation of NF-κB pathway

目的探究DDR1促进胃癌细胞迁移侵袭的作用机制。方法通过慢病毒转染法构建SNU719细胞DDR1过表达稳转株,嘌呤霉素药筛后使用RT-qPCR和蛋白印迹法进行稳转效果验证。利用Transwell小室实验对比DDR1过表达细胞株和对照组细胞的迁移侵袭能力,通过Western Blot和抑制剂探究DDR1过表达对SNU719下游信号通路的影响。结果RT-qPCR和蛋白印迹结果显示在SNU719过表达组细胞株中,DDR1在mRNA水平和蛋白水平均有明显的升高。迁移侵袭实验示,小室膜底面的过表达组SNU719细胞数量均显著高于对照组。Western Blot结果表明过表达DDR1组SNU719核内B-catenin含量明显增加,而NF-κB抑制剂亦明显抑制DDR1过表达组细胞的迁移侵袭能力。结论DDR1通过上调NF-κB通路促进胃癌细胞的迁移侵袭。

Objective To explore the mechanism underlying that DDR1 promotes the metastasis ability of gastric cancer cell line SNU719.Methods The DDR1 overexpression stable strain of SNU719 cells was constructed by lentiviral transfection and verified by RT-PCR and Western blotting after puromycin selection.Transwell chamber assay was used to compare the migration and invasion abilities of DDR1 overexpressing cells and control cells.The effects of DDR1 overexpression on the downstream signaling pathway of SNU719 were determined by Western Blot and inhibitors.Results RT-PCR and Western blot showed that DDR1 significantly increased in mRNA and protein levels in SNU719 overexpressing cell line.The migration and invasion assay showed that the number of SNU719 cells in the DDR1 group on the bottom of the small chamber membrane was significantly higher than that in the control group.The results of Western blot suggested that the content of b-catenin in SNU719 nucleus of over expressed DDR1 group increased significantly,and NF-kB inhibitor also decreased the migration and invasion ability of cells in over expressed DDR1 group.Conclusions DDR 1 promotes the migration and invasion of gastric cancer cells by upregulating the NF-κB pathway.