LncRNA PTCSC3过表达诱导非小细胞肺癌A549凋亡和氧化损伤并抑制细胞增殖和迁移

Overexpression of LncRNA PTCSC3 induces apoptosis and oxidative damage of non-small cell lung cancer A549 and inhibits their proliferation and migration

目的:探究LncRNA PTCSC3过表达对非小细胞肺癌A549的影响。方法:将A549细胞分为对照组、pcDNA组和pcDNA-lncRNA PTCSC3组,采用聚合酶链反应检测lnc PTCSC3、Ki67和p21mRNA表达,流式细胞仪检测细胞凋亡,试剂盒检测活性氧簇含量,克隆形成检测细胞增殖,Transwell小室实验检测细胞侵袭,蛋白印迹法检测凋亡(caspase-3和caspase-9)和侵袭(E-cadherin和N-cadherin)相关蛋白的表达。结果:与pcDNA组相比,pcDNA-lncRNA PTCSC3组PTCSC3基因、细胞凋亡率、cleaved caspase-3/caspase-3、cleaved caspase-9/caspase-9、p21 mRNA及E-cadherin蛋白水平均显著升高(t=14.758、12.188、9.905、7.320、13.975、7.443,均P<0.05),活性氧簇含量、克隆形成率、Ki67 mRNA、单位面积侵袭细胞数目及N-cadherin蛋白水平均显著降低(t=10.276、9.615、20.444、6.576、14.992,均P<0.05)。结论:过表达lncRNA PTCSC3通过调控氧化应激和相关蛋白表达水平,抑制非小细胞肺癌A549的增殖和迁移并促进其凋亡。

Objective:To explore the effects of LncRNA PTCSC3 overexpression on non-small cell lung cancer(NSCLC)A549.Methods:A549 cells were divided into control group,pcDNA group and pcDNA-lncRNA PTCSC3 group.The expression of lnc PTCSC3,Ki67 and p21mRNA was detected by polymerase chain reaction.The apoptosis was detected by flow cytometry.The contents of reactive oxygen species(ROS)were detected by kits.The proliferation of cells was detected by clone formation assay.The invasion of cells was detected by Transwell chamber assay.The expression of apoptosis related proteins(caspase-3,caspase-9)and invasion related proteins(E-cadherin,N-cadherin)were detected by Western blotting.Results:Compared with pcDNA group,PTCSC3 gene,apoptosis rate,levels of cleaved caspase-3/caspase-3,cleaved caspase-9/caspase-9,p21mRNA and E-cadherin protein in pcDNA-lncRNA PTCSC3 group were significantly increased(t=14.758,12.188,9.905,7.320,13.975,7.443,all P<0.05),while contents of ROS,clone formation rate,Ki67 mRNA,number of invasion cells per unit area and level of N-cadherin protein were significantly decreased(t=10.276,9.615,20.444,6.576,14.992,all P<0.05).Conclusion:LncRNA PTCSC3 overexpression inhibits the proliferation and migration of NSCLC A549,and promotes its apoptosis by regulating oxidative stress and expression levels of related proteins.