NFATc1对非小细胞肺癌细胞增殖、迁移和上皮间质转化的影响

Effects of NFATc1 on the proliferation,migration and epithelial-mesenchymal transition of non-small cell lung cancer

目的:检测活化T细胞c1核因子(NFATc1)对非小细胞肺癌(NSCLC)细胞增殖、迁移和上皮间充质转化(EMT)的影响。方法:qRT-PCR检测NSCLC细胞H1650、H460、H1299及正常人支气管上皮细胞BEAS-2B中NFATc1表达水平,将si-NFATc质粒转染至H1650细胞中,通过CCK-8增殖及集落形成实验、划痕和Western印迹实验分别检测转染后H1650细胞增殖、迁移及EMT的影响。结果:NFATc1 mRNA在H1650、H460、H1299细胞中的相对表达水平高于BEAS-2B细胞(t=10.732,P<0.001;t=4.705,P=0.009;t=4.792,P=0.009),且H1650细胞中NFATc1 mRNA表达水平高于H460、H1299细胞(t=3.593,P=0.023;t=7.505,P=0.002);转染si-NFATc1组NFATc1 mRNA(t=9.325,P<0.001),NFATc1蛋白(t=10.254,P<0.001),细胞的OD值(t=7.954,P<0.001)均低于si-NC组,细胞集落形成数(t=12.210,P<0.001),迁移率(t=8.951,P<0.001)低于si-NC组;si-NFATc1组E-cadherin蛋白表达水平高于si-NC组(t=6.352,P<0.001),Vimentin和N-cadherin蛋白的表达(t=6.012,P<0.001;t=10.241,P<0.001)低于si-NC组。结论:NFATc1在H1650细胞中表达水平增加,敲低其表达水平后可减弱细胞增殖、迁移和EMT。

Objective:To detect the effect of activated T cell c1 nuclear factor(NFATc1)on the proliferation,migration and epithelial-mesenchymal transition(EMT)of non-small cell lung cancer(NSCLC)cells.Methods:qRT-PCR was used to detect the expression level of NFATc1 in NSCLC cells H1650,H460,H1299 and normal human bronchial epithelial cells BEAS-2B.The si-NFATc plasmid was transfected into H1650 cells,and the effects of H1650 cell proliferation,migration and epithelial-mesenchymal transition(EMT)were detected by CCK-8 proliferation,colony formation experiments,scratch and Western blotting.Results:The relative expression level of NFATc1 mRNA in H1650,H460,and H1299 cells were higher than those in BEAS-2B cells(t=10.732,P<0.001;t=4.705,P=0.009;t=4.792,P=0.009),and the expression level of NFATc1 mRNA of H1650 cells was higher than that of H460 and H1299 cells(t=3.593,P=0.023;t=7.505,P=0.002).The NFATc1 mRNA(t=9.325,P<0.001),and the gray value of NFATc1 protein(t=10.254,P<0.001),the OD value of cells(t=7.954,P<0.001)in the si-NFATc1 group were lower than those of the si-NC group,the number of colony formation(t=12.210,P<0.001),and the migration rate in the si-NFATc1 group were lower than those of the si-NC group(t=8.951,P<0.001).The expression of E-cadherin protein in the si-NFATc1 group was higher(t=6.352,P<0.001),and the expression of Vimentin and N-cadherin protein were lower than those in the si-NC group(t=6.012,P<0.001;t=10.241,P<0.001).Conclusion:The expression level of NFATc1 increases in H1650 cells,knocking down its expression level can attenuate cell proliferation,migration and EMT.