circTADA2A在肺腺癌组织和细胞中的表达及其对A549细胞迁移及侵袭影响的实验研究

Expression of circTADA2A in lung adenocarcinoma tissues and cells and its effect on A549 cells migration and invasion

目的:研究环状RNA TADA2A(circular RNA TADA2A,circTADA2A)在肺腺癌(lung adenocarcinoma,LAD)组织和细胞中的表达及其对A549细胞迁移及侵袭的影响。方法:在Gene Expression Omnibus(GEO)数据库中下载非编码RNA阵列分析文件GSE101586,并通过在线分析软件GEO2R分析GSE101586中5例肺腺癌及配对的癌旁组织中差异表达的环状RNA(circular RNAs,circRNAs);筛选差异表达的circRNAs并绘制热图;检测circTADA2A在GSE101586的5例肺腺癌及配对的癌旁组织中的差异性表达;qRT-PCR法检测circTADA2A在非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞系A549及人支气管上皮细胞系16HBE中的差异表达;在A549细胞中转染特异性circTADA2A干扰寡核苷酸(circTADA2A small interfering RNA,sicircTADA2A),同步转染错义对照寡核苷酸(small interfering scramble,siSCR),qRT-PCR法检测circTADA2A的差异表达;Transwell迁移及侵袭实验检测不同circTADA2A表达水平对A549细胞迁移及侵袭能力的影响。结果:在GSE101586中,相比于癌旁组织,在肺腺癌组织中差异表达显著的circRNAs有39个(筛选标准:P<0.05且|logFC|≥1.2),其中上调的circRNAs有22个,下调的circRNAs有17个;相比于癌旁组织,circTADA2A在肺腺癌组织中表达升高;相比于16HBE细胞,circTADA2A在A549细胞中表达升高;转染sicircTADA2A可明显敲低A549细胞内circTADA2A的表达水平;下调circTADA2A的表达水平可明显抑制A549细胞的迁移及侵袭能力。结论:circTADA2A在肺腺癌组织和细胞中表达增高,下调其表达可明显抑制A549细胞的迁移及侵袭能力。

Objective:To explore the expression of circular RNA TADA2A(circTADA2A)in lung adenocarcinoma(LAD)tissues and cells,and is to evaluate the effect of circTADA2A on A549 cells migration and invasion.Methods:The differentially expressed circRNAs data of 5 LAD tissues and paired paratumor tissues in GSE101586 was downloaded from Gene Expression Omnibus(GEO)database.The data was analyzed by online software GEO2R.Expression of circTADA2A in GSE101586 was detected by GEO2R,either.Expression of circTADA2A in a non-small cell lung cancer cell line A549 and in a human normal bronchial epithelial cell line 16HBE was detected by qRT-PCR.Specific small interfering RNA(siRNA)targeted circTADA2A(sicircTADA2A)and corresponding scramble siRNA(siSCR)were transfected into A549 cells,followed by a qualification of circTADA2A via qRT-PCR.The effect of different level of circTADA2A on cell migration and invasion were determined by Transwell assay in A549 cells.Results:A total of 39 circRNAs were differentially expressed in GSE101586 between LAD and paratumor tissue(filtration criteria:P<0.05 and|LogFC|value≥1.2).Among which,22 circRNAs were upregulated,and 17 circRNAs were downregulated.circTADA2A was upregulated in LAD tissues as comparing with that in paratumor tissues.Meanwhile,circTADA2A was upregulated in A549 cells as comparing with that in 16HBE.A transfection of sicircTADA2A suppressed the level of circTADA2A in A549 cells.A downregulation of circTADA2A inhibited migration and invasion ability of A549 cells.Conclusion:circTADA2A was upregulated in LAD,and a suppression of circTADA2A inhibited migration and invasion ability of A549 cells.