长链非编码RNA MALAT1对NSCLS增殖、侵袭迁移及凋亡的影响

Effects of long non-coding RNA MALAT1 on proliferation,invasion,migration and apoptosis of NSCLS

目的探究长链非编码RNA(lncRNA)肺腺癌转移相关转录物1(MALAT1)对非小细胞型肺癌(NSCLC)增殖、侵袭迁移及凋亡的影响及其作用机制。方法收集2015年6月至2017年8月在天门市第一人民医院接受肺癌手术的34例NSCLC患者的肿瘤组织和癌旁正常组织标本。体外培养肺癌A549细胞,并分为空白对照(Control)组、阴性对照(shRNA-NC)组和敲除lncRNA MALAT1(sh-MALAT1)组;shRNA-NC组使用shRNA-NC转染肺癌A549细胞;sh-MALAT1组利用干扰RNA(siRNA)敲除lncRNA MALAT1并转染肺癌A549细胞。采用实时定量PCR(RT-PCR)法检测MALAT1的表达情况;克隆形成实验检测肺癌A549细胞生长情况;流式细胞仪检测细胞凋亡情况;Transwell检测细胞侵袭情况;Western blot检测细胞中增殖细胞周期相关核抗原(Ki67)、Caspase-9、E-钙黏蛋白(E-cadherin)及N-钙黏蛋白(N-cadherin)的表达情况。结果肿瘤组织中MALAT1表达水平显著高于正常组织(P<0.05);与Control组相比,shRNA-NC组MALAT1表达水平、细胞克隆形成率、细胞凋亡率、细胞侵袭情况均无显著变化(P>0.05);与shRNA-NC组相比,sh-MALAT1组MALAT1表达水平、细胞克隆形成率、单位面积细胞侵袭数目Ki67和N-cadherin蛋白表达水平显著降低(P<0.05),细胞凋亡率、Caspase-9和E-cadherin蛋白表达水平显著升高(P<0.05)。结论lncRNA MALAT1在肺癌组织中过表达,敲低NSCLC细胞中MALAT1的表达水平可以抑制NSCLC细胞的增殖、侵袭及迁移并促进NSCLC细胞的凋亡。

Objective To explore effects of long non-coding RNA(lncRNA)MALAT1 on proliferation,invasion,migration and apoptosis of non-small cell lung cancer(NSCLC)and its action mechanism.Methods Tumor samples and adjacent normal tissues of 34 NSCLC patients who underwent lung cancer surgery in Oncology Department of Tianmen First People's Hospital were collected.Lung cancer A549 cells were cultured in vitro.And they were divided into blank control group(control),negative control group(shRNA-NC)and knockout lncRNA MALAT1 group(sh-MALAT1).In shRNA-NC group,lung cancer A549 cells were transfected with shRNA-NC.In sh-MALAT1 group,lncRNA MALAT1 was knocked out by small interference RNA(siRNA)and lung cancer A549 cells were transfected.The expression of MALAT1 was detected by real-time PCR(RT-PCR).The growth of lung cancer A549 cells was detected by colony formation assay.The apoptosis of lung cancer A549 cells was detected by flow cytometry.Transwell was applied to detect cell invasion.The expression of Ki67,Caspase-9,E-cadherin and N-cadherin protein in cells was detected by Western blot.Results The expression level of MALAT1 in tumor tissues was significantly higher than that in normal tissues(P<0.05).Compared with control group,there was no significant change in expression level of MALAT1,cell clone formation rate,apoptosis rate or cell invasion in shRNA-NC group(P>0.05).Compared with shRNA-NC group,MALAT1 expression level,cell clone formation rate,number of cell invasion per unit area,expression levels of Ki67 and N-cadherin in sh-MALAT1 group were significantly decreased(P<0.05),while apoptosis rate,expression levels of Caspase-9 and E-cadherin protein were significantly increased(P<0.05).Conclusions lncRNA MALAT1 is over-expressed in lung cancer tissues.Knocking down expression level of MALAT1 in NSCLC cells can inhibit proliferation,invasion and migration of NSCLC cells,and promote apoptosis of NSCLC cells.