敲低WTAP基因对肺腺癌A549细胞恶性生物学行为的影响

Effect of WTAP gene knockdown on malignant biological behaviors of lung adenocarcinoma A549 cells

目的:探讨成肾细胞瘤1-结合蛋白(Wilms'tumor 1-associating protein,WTAP)对人肺腺癌A549细胞增殖、迁移和侵袭的影响。方法:选用人肺腺癌A549细胞和HEK293T,设计两组shWTAP干扰序列,构建慢病毒载体质粒,在HEK293T细胞中包装慢病毒后感染人肺腺癌A549细胞,对照组感染277空载体质粒。用qPCR和WB实验检测A549细胞中WTAP mRNA和蛋白的表达水平,用BrdU实验、细胞划痕愈合实验、Transwell实验分别检测A549细胞的增殖、迁移和侵袭能力。结果:成功构建shWTAP-1和shWTAP-2两种慢病毒质粒,感染A549细胞后,与对照组比较,A549细胞中WTAP mRNA和蛋白表达水平显著下调(均P<0.05),细胞的增殖、迁移和侵袭能力显著降低(均P<0.05)。结论 :敲低WTAP基因的表达对人肺腺癌A549细胞的增殖、迁移和侵袭能力有显著抑制作用,WTAP基因的表达与肺腺癌的发生发展相关,WTAP可能是肺腺癌诊疗的一个潜在靶标。

Objective:To investigate the effects of Wilms’tumor 1-associating protein(WTAP)on proliferation,migration and invasion of human lung adenocarcinoma A549 cells.Methods:Human lung adenocarcinoma cell line A549 and HEK293 T cells were chosen for this study.Two sets of shWTAP interference sequences were designed to construct lentiviral vector plasmid.Human lung adenocarcinoma A549 cells were infected after packaging lentivirus in HEK293 T cells,and the control group was transfected with 277 empty vector plasmid.The mRNA and protein expression levels of WTAP in A549 cells were detected by qPCR and WB.Changes in proliferation,migration and invasion of A549 cells were detected by BrdU assay,cell scratch healing assay and Transwell assay,respectively.Results:Two plasmids,shWTAP-1 and shWTAP-2,were successfully constructed.Compared with the control group,the mRNA and protein expression levels of WTAP were significantly down-regulated in A549 cells with WTAP knockdown(both P<0.05),and the proliferation,migration and invasion ability of cells were significantly decreased(all P<0.05).Conclusion:Knockdown of WTAP significantly inhibited the proliferation,migration and invasion of human lung adenocarcinoma A549 cells.The expression of WTAP gene is associated with the occurrence and development of lung adenocarcinoma.WTAP may be a potential target for the diagnosis and treatment of lung adenocarcinoma.