NFATc1对人肺腺癌细胞增殖、凋亡和侵袭能力的影响

Effects of NFATc1 on the proliferation,apoptosis and invasion of human lung adenocarcinoma cells

目的探究NFATc1对肺腺癌NCI-H1299细胞增殖、凋亡、迁移和侵袭能力的影响,以及NFATc1发挥作用的生物学机制。方法通过Realtime PCR检测NFATc1在三种肺腺癌细胞系中的相对表达水平,利用慢病毒载体构建稳定敲减NFATc1的NCI-H1299细胞系。MTT检测NFATc1对细胞增殖能力的影响,平板克隆形成实验检测NFATc1对克隆形成能力的影响,Transwell检测NFATc1对细胞迁移和侵袭能力的影响,流式细胞术检测NFATc1对细胞周期和凋亡的影响,Western blot检测NFATc1对凋亡、EMT相关蛋白和MAPK信号通路蛋白表达的影响。结果肺腺癌NCI-H1299细胞系中NFATc1表达水平相对较高。沉默NFATc1表达可以显著抑制细胞增殖(P<0.05)、克隆形成(P<0.05)、迁移(P<0.05)和侵袭(P<0.05)能力,抑制细胞周期在G1期(P<0.05),促进细胞凋亡(P<0.05)。Bax、Cleaved caspase-3和E-Cadherin蛋白表达增加(P<0.05),CDK4、c-Myc、ERK、p-ERK和N-Cadherin蛋白表达降低(P<0.05)。结论在人肺腺癌NCI-H1299细胞中抑制NFATc1表达,可以显著抑制细胞增殖、细胞周期、迁移和侵袭,并促进细胞凋亡,其机制可能与抑制MAPK信号通路相关。

Objective The aim of this study was to investigate the effects of NFATc1 expression on the proliferation,apoptosis,migration and invasion of lung adenocarcinoma NCI-H1299 cells,and its specific biological mechanisms.Methods Real time PCR was used to detect the relative levels of NFATc1 in three lung adenocarcinoma cell lines,and the NCI-H1299 cell line with stable knockdown of NFATc1 was constructed using lentivirus vectors.A MTT assay was used to detect the effect of NFATc1 on cell proliferation,the cell colony assay was used to detect the effect of NFATc1 on clone formation,the Transwell assay was used to detect the effect of NFATc1 on cell migration and invasion,and flow cytometry was used to detect the effect of NFATc1 on cell cycle and apoptosis.Western blot was used to detect the effects of NFATc1 on apoptosis,EMT-related proteins and MAPK signaling pathways protein expression.Results The expression of NFATc1 in lung adenocarcinoma NCI-H1299 cells was relatively high.Silencing NFATc1 expression could significantly inhibit cell proliferation(P<0.05),clone formation(P<0.05),migration(P<0.05)and invasion(P<0.05),the cell cycle arrested at the G1 phase(P<0.05),and promote apoptosis(P<0.05).The expression of Bax,cleaved caspase-3 and E-Cadherin protein increased(P<0.05),and CDK4,c-Myc,ERK,p-ERK and N-Cadherin protein expression decreased(P<0.05).Conclusion Inhibition of NFATc1 expression in human lung adenocarcinoma NCI-H1299 cells can significantly inhibit cell proliferation,arresting cell cycle,migration and invasion,and promote cell apoptosis.The mechanism may be related to the inhibition of MAPK signaling pathway.