YAP对非小细胞肺癌细胞增殖和迁移能力的影响

Effect of YAP on proliferation and migration of non-small cell lung cancer cells

目的:探讨长链非编码RNA(lncRNA)MALAT1在YAP调控的非小细胞肺癌(NSCLC)细胞增殖和迁移中的作用,并阐明其作用机制。方法:选择A549和H1299细胞系分为Scramble siRNA组(转染Scramble siRNA)、siYAP-1组(转染siYAP-1)和siYAP-2组(转染siYAP-2)。RT-PCR和Western blotting法检测各组细胞中YAP和MALAT1 mRNA及YAP蛋白表达水平;在A549和H1299细胞中分别转染pcDNA3.1-YAP质粒(pcDNA3.1-YAP组)、共转染pcDNA3.1-YAP质粒和siMALAT1-2(pcDNA3.1-YAP+siMALAT1-2组)及对照质粒(pcDNA3.1-YAP+Scramble siRNA组),CCK-8法检测各组细胞增殖能力,细胞划痕实验检测各组细胞迁移能力。结果:siYAP-1组和siYAP-2组A549和H1299细胞中YAP mRNA及蛋白表达水平均明显低于Scramble siRNA组(P<0.05),siYAP-1组和siYAP-2组A549和H1299细胞中MALAT1 mRNA表达水平低于Scramble siRNA组(P<0.05);与pcDNA3.1-YAP组比较,pcDNA3.1-YAP+siMALAT1-2组A549和H1299细胞增殖能力降低(P<0.05),H1299细胞迁移能力降低(P<0.05)。结论:在NSCLC细胞中,YAP通过调控lncRNA MALAT1的表达促进细胞的增殖和迁移。

Objective:To investigate the role of long non-coding RNA(lncRNA)MALAT1 in theProliferation and migration of YAP-mediated non-small cell lung cancer(NSCLC)cells,and to elucidate its mechanism.Methods:The A549 and H1299 cell lines were selected and divided into Scramble siRNA group(transfected with Scramble siRNA),siYAP-1 group(transfected with siYAP-1)and siYAP-2 group(transfected with siYAP-2).RT-PCR and Western blotting methods were used to detect the expression levels of YAP and MALAT1 mRNA and YAPProtein in the cells in variouis groups.ThePcDNA3.1-YAPPlasmid(pcDNA3.1-YAP group),PcDNA3.1-YAPPlasmidPlus siMALAT1-2(pcDNA3.1-YAP+siMALAT1-2 group)and the controlPlasmid(pcDNA3.1-YAP+Scramble siRNA group)were transfected into the A549 and H1299 cells,respectively.CCK-8 method and scratch assay were used to detect theProliferation and migration abilities of the cells in various groups.Results:The expression levels of YAP mRNA andProtein in the A549 and H1299 cells in siYAP-1 and siYAP-2 groups were significantly lower than those in Scramble siRNA group(P<0.05),and the expression levels of MALAT1 mRNA in the cells in siYAP-1 and siYAP-2 groups were significantly lower than those in Scramble siRNA group(P<0.05).Compared withPcDNA3.1-YAP group,theProliferation abilities of the A549 and H1299 cells inPcDNA3.1-YAP+siMALAT1-2 group were decreased(P<0.05),the migration ability of the H1299 cells inPcDNA3.1-YAP+siMALAT1-2 group was decreased(P<0.05).Conclusion:In the NSCLC cells,YAPPromotes the cellProliferation and migration by regulating the expression of lncRNA MALAT1.