LINC01176对非小细胞肺癌细胞侵袭和迁移的影响及其与上皮-间质转化的关系

Effects of LINC01176 on invasion and migration of non-small-cell lung cancer cells and their relation⁃ship with epithelial-mesenchymal transition

目的观察长链非编码RNA LINC01176对非小细胞肺癌(NSCLC)细胞侵袭和迁移的影响,并分析该作用是否与上皮—间质转化有关。方法选取人NSCLC细胞A549、H1299、H1975、H1650和正常肺上皮细胞16HBE,采用实时荧光定量PCR法检测各细胞中LINC01176的表达水平。取LINC01176表达最高的NSCLC细胞分为转染敲低组和转染对照组,分别转染LINC01176 siRNA和LINC01176 siRNA对照无干扰序列。采用Transwell实验观察细胞侵袭能力,细胞划痕实验观察细胞迁移能力。通过实时荧光定量PCR法和Western Blotting法分别检测细胞上皮—间质转化相关标志物E-cadherin、Vimentin的mRNA和蛋白相对表达量。结果LINC01176在NSCLC细胞中的相对表达量均高于正常肺上皮细胞(P均<0.05),其中H1975细胞的LINC01176表达量最高;转染敲低组穿膜细胞数、细胞迁移距离均小于转染对照组(P均<0.05);转染敲低组E-cadherin mRNA及蛋白表达量均高于转染对照组,Vimentin mRNA及蛋白表达量均低于转染对照组(P均<0.05)。结论LINC01176高表达可促进NSCLC细胞的侵袭和迁移,其作用可能通过参与调控上皮—间质转化实现。

Objective To investigate the effects of long non-coding RNA(LncRNA)LINC01176 on the cell invasion and migration of non-small-cell lung cancer(NSCLC)and to analyze whether the effects were related to the epithelial-mesenchymal transition(EMT).Methods The expression levels of LINC01176 in NSCLC cells(A549,H1299,H1975 and H1650)and normal lung epithelial cells(16HBE)was detected by real-time fluorescent quantitative PCR(RT-qPCR).H1975 cells with the highest expression of LINC01176 were divided into two groups:the LINC01176 knockdown group which was transfected with the LINC01176 siRNA,and the negative control group which was transfected with the control siRNA.The cell invasion ability was observed by Transwell assay,and the cell migration ability was observed by Scratch assay.The relative mRNA and protein expression levels of EMT-related markers(E-cadherin and Vimentin)were detected by RT-qPCR and Western blotting.Results The expression levels of LINC01176 in NSCLC cells(A549,H1299,H1975 and H1650)were higher than that in normal lung epithelial cells(16HBE),and the expression level of LINC01176 in H1975 cells was the highest(all P<0.05).In H1975 cells,the number of transmembrane cells and migra⁃tion distance in the LINC01176 knockdown group were lower than those in the negative control group(both P<0.05),the mRNA and protein expression levels of E-cadherin in the LINC01176 knockdown group were higher than those in the negative control group,and the mRNA and protein expression levels of Vimentin in the LINC01176 knockdown group were lower than those in the negative control group(all P<0.05).Conclusion High expression of LINC01176 can promote the invasion and migration of NSCLC cells by participating in the regulation of EMT.