长链非编码RNA ZFAS1在非小细胞肺癌患者组织中表达及作用

摘要

目的 检测非小细胞肺癌(NSCLC)患者癌组织中锌指结构反义转录本1(zinc finger antisense 1,ZFAS1)表达水平,探讨ZFASI在NSCLC进展中的生物学作用.方法 实时荧光定量RT-PCR检测ZFAS1在NSCLC患者癌组织和癌旁组织中的表达水平.RNA干扰ZFAS1在A549细胞中表达,细胞计数和克隆形成实验检测细胞增殖情况,流式分析检测细胞周期和细胞凋亡,Transwell迁移和基质胶侵袭实验检测细胞迁移和侵袭能力,实时荧光定量RT-PCR检测Cyclin D1、Bcl2、N-cadherin、ZEB1、Slug和Twist基因表达水平变化.结果 ZFAS1在NSCLC患者癌组织中的平均表达水平[0.01 (0.002,0.054)]较癌旁组织[0.002 (0.001,0.012)]明显升高(Z=-2.638,P＜0.01).ZFAS1基因敲减后,A549细胞增殖能力明显减弱(P＜0.01);A549细胞周期G1期比例升高,S期比例下降(P ＜0.01);A549细胞凋亡比例明显增加(P ＜0.01);A549细胞迁移和侵袭能力明显下降(P均＜0.01);A549细胞中Cyclin D1、Bcl2、N-cadherin、ZEB1、Slug和Twist基因表达水平均降低(P均＜0.05).结论 ZFAS1在NSCLC患者癌组织中呈高表达.ZFAS1基因敲减诱导细胞周期阻滞、凋亡和抑制上皮-间质转变(EMT),减弱NSCLC细胞增殖、迁移和侵袭能力.%Objective To determine the expression levels of zinc finger antisense 1 (ZFAS1) in tumor tissues of non-small cell lung cancer (NSCLC) and investigate the biological roles of ZFAS1 in NSCLC.Methods The relative expression levels of ZFAS1 in tumor tissues of NSCLC patients were determined by using real-time fluorescent qRT-PCR.RNA interference was used to knock down ZFAS1 expression in A549 cells.The proliferations of A549 cells with ZFAS1 knockdown were determined by cell counting and cell colony formation assays.Flow cytometric analysis was used to determine the cellular cycle distribution and apoptosis of the A549 cells with ZFAS1 knockdown.Transwell migration and matrigel invasion assays were used to determine the migration and invasion of the A549 cells with ZFAS1 knockdown.The gene expression levels of cyclin D1,Bcl2,N-cadherin,ZEB1,Slug and Twist were determined by real-time fluorescent qRT-PCR.Results The mean expression levels of ZFAS1 in tumor tissues of NSCLC patients [0.01 (0.002 to 0.054)] were significantly higher than those in the adjacent non-cancerous tissues [0.002 (0.001 to 0.012)] (Z =-2.638;P ＜ 0.01).After ZFAS1 knockdown,the proliferation of A549 cells was remarkably retarded (P ＜ 0.01).The percentage of cells at G1 phase was increased while the cells at S phase was decreased (P ＜ 0.01).The rate of apoptotic cells was significantly increased in A549 cells with ZFAS1 knockdown (P ＜ 0.01).A549 cells showed decreased migration and invasion abilities after ZFAS1 knockdown (P ＜0.01).The expression levels of cyclin D1,Bcl2,N-cadherin,ZEB1,Slug and Twist genes were decreased in ZFAS1 knockdown A549 cells (P ＜ 0.05).Conclusion ZFASI was highly expressed in the tumor tissues of NSCLC patients.ZFAS1 knockdown induced cell cycle arrest,cell apoptosis and suppression of epithelial-mesenchymal transition (EMT),leading to the inhibition of proliferation,migration,and invasion of NSCLC cells.