长链非编码RNA PRNCR1对非小细胞肺癌A549细胞迁移和凋亡的影响

Effects of long chain non-coding RNA PRNCR1 on migration and apoptosis of A549 cells in non-small cell lung cancer

目的探究长链非编码RNA PRNCR1对非小细胞肺癌A549细胞迁移和凋亡的影响。方法构建shRNA lncRNA PRNCR1慢病毒载体,以人肺腺癌细胞系A549为试验材料,将细胞分成三组,即空白对照组,阴性对照组和实验组。空白对照组细胞未做任何处理,阴性对照组是空白载体慢病毒转染的细胞,实验组是由shRNA lncRNA PRNCR1的慢病毒干扰载体构建的慢病毒转染细胞。RT-PCR和Western blot检测lncRNA PRNCR1,以及凋亡因子Caspase-3和Caspase-9的表达,Transwell试剂盒检测细胞的迁移,AV-PI试剂盒检测细胞的凋亡。结果shRNA-lncRNA PRNCR1载体慢病毒转染滴度为3×10 8 TU/ml。RT-PCR结果显示,实验组lncRNA PRNCR1的mRNA的表达水平明显低于阴性对照组,差异有统计学意义(P<0.05),实验组Caspase-3、Caspase-9的mRNA水平均明显高于阴性对照组,差异有统计学意义(均P<0.05)。Western blot检测结果和RT-PCR结果相同。Transwell检测结果显示,实验组与阴性对照组相比,非小细胞癌A549细胞的迁移率明显降低,差异具有统计学意义(P<0.05);AV-PI检测结果显示,实验组与阴性对照组相比,非小细胞癌A549细胞的凋亡率明显升高,差异具有统计学意义(P<0.05)。结论shRNA lncRNA PRNCR1可以降低非小细胞肺癌A549细胞的迁移率,提高凋亡率,为非小细胞肺癌的治疗提供思路。

Objective To investigate the effect of long-stranded non-coding RNA(lncRNA)PRNCR1 on migration and apoptosis of non-small cell lung cancer A549 cells.Methods The shRNA-lncRNA PRNCR1 lentiviral vector was constructed in this study.Human lung adenocarcinoma cell line A549 cells were used as the experimental material.The cells were divided into three groups:blank control group,negative control group and experimental group.The cells in blank control group were not treated,the cells in negative control group were transfected with blank vector by lentiviral virus,and the cells in experimental group were transfected with shRNA lncRNA PRNCR1 interference vector by lentiviral virus.The expression levels of lncRNA PRNCR1,apoptotic factors Caspase-3 and Caspase-9 were detected by RT-PCR and Western blot,the migration of cells was detected by Transwell kit,and the apoptosis was detected by AV-PI kit.Results The lentiviral titer of shRNA-lncRNA PRNCR1 vector was 3×10 8 TU/ml.RT-PCR results showed that the expression level of lncRNA PRNCR1 in experimental group was significantly lower than that in negative control group(P<0.05).The expression levels of Caspase-3 and Caspase-9 mRNA in experimental group were significantly higher than those in negative control group(P<0.05).The Western blot results were the same as that of RT-PCR.The results of Transwell assay showed that the migration rate of non-small cell carcinoma A549 cells was significantly lower in experiment group than in negative control group(P<0.05),and the results of AV-PI kit showed that the migration rate of non-small cell carcinoma A549 cells was significantly lower in experiment group than in negative control group(P<0.05).Conclusion The shRNA lncRNA PRNCR1 can reduce the migration rate of non-small cell lung cancer A549 cells,improve the rate of apoptosis,and provide ideas for clinical treatment of non-small cell lung cancer.