LncRNA TUG1调控miR-142-5p/PD-L1轴促进非小细胞肺癌细胞增殖、迁移与侵袭能力

LncRNA TUG1 promotes cell proliferation,migration,and invasion through regulating the miR-142-5p/PD-L1 axis in non-small cell lung cancer cells

目的探讨长链非编码RNA(long non-coding RNA,LncRNA)牛磺酸上调基因1(taurine up-regulated gene 1,TUG1)对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞生物学行为的影响及分子机制。方法采用qRT-PCR、Western blot分别检测组织(NSCLC癌组织、癌旁组织)和细胞(人正常气管上皮细胞HBE及NSCLC细胞A549、H2009、H1975)中TUG1、miR-142-5p及PD-L1的表达。将pcDNA-TUG1、si-TUG1、miR-142-5p mimics、si-TUG1+anti-miR-142-5p分别转染于A549,qRT-PCR、Western blot分别检测细胞中TUG1、miR-142-5p及PD-L1表达;CCK-8检测细胞增殖;Transwell检测细胞迁移、侵袭;双荧光素酶基因实验分别检测TUG1和miR-142-5p、miR-142-5p和PD-L1的关系;RNA免疫共沉淀检测miR-142-5p与TUG1的相互作用。结果NSCLC组织中TUG1(2.28±0.23 vs 1.00±0)、PD-L1(0.93±0.11 vs 0.25±0.01)表达高于癌旁组织,miR-142-5p(0.31±0.02 vs 1.00±0)表达低于癌旁组织(P<0.05);与人正常气管上皮细胞HBE比较,NSCLC细胞A549中TUG1(3.21±0.27 vs 1.00±0)、PD-L1(1.12±0.12 vs 0.24±0.01)表达水平最高,miR-142-5p(0.23±0.02 vs 1.00±0)表达水平最低(P<0.05);过表达TUG1促进A549细胞增殖、迁移、侵袭;下调TUG1或上调miR-142-5p表达均对A549细胞增殖、迁移、侵袭能力发挥抑制作用;TUG1负调控miR-142-5p,miR-142-5p负调控PD-L1;下调miR-142-5p减弱了沉默TUG1对A549细胞增殖、迁移及侵袭的抑制作用。结论TUG1在NSCLC组织和细胞中高表达,沉默TUG1通过miR-142-5p/PD-L1轴抑制A549细胞的增殖、迁移与侵袭。

Purpose To investigate the effects of long non-coding RNA taurine up-regulated gene 1(LncRNA TUG1)on cell proliferation,migration and invasion of non-small cell lung cancer(NSCLC)and its molecular mechanism.Methods RT-qPCR and Western blot were used to detect TUG1,miR-142-5p and PD-L1 protein expression in tissues(NSCLC cancer tissues,adjacent tissues)and cells(human normal tracheal epithelial cells HBE and NSCLC cells A549,H2009,H1975),respectively.A549 was transfected with pcDNA-TUG1,si-TUG1,miR-142-5p mimics,and si-TUG1+anti-miR-142-5p,respectively.The expressions of TUG1,miR-142-5p and PD-L1 protein in cells were detected by qRT-PCR and Western blot,respectively.CCK-8 was used to detect cell proliferation.Transwell was used to detect cell migration and invasion,dual luciferase gene assay was to detect the relationship between TUG1 and miR-142-5p,miR-142-5p and PD-L1,respectively.Inter-action between miR-142-5p and TUG1 was detected by RNA co-immunoprecipitation(RIP).Results Compared with adjacent tissues,the TUG1(2.28±0.23 vs 1.00±0)and PD-L1 protein(0.93±0.11 vs 0.25±0.01)expression levels in NSCLC tissues were higher,and the miR-142-5p(0.31±0.02 vs 1.00±0)expression level was lower(P<0.05).Compared with human normal tracheal epithelial cells HBE,the expression levels of TUG1(3.21±0.27 vs 1.00±0)and PD-L1 protein(1.12±0.12 vs 0.24±0.01)were the highest,and miR-142-5p(0.23±0.02 vs 1.00±0)with the lowest expression level in NSCLC cells A549(P<0.05).Overexpression of TUG1 promoted the proliferation,migration and invasion of A549 cells,downregulating of TUG1 or upregulating of miR-142-5p could inhibit the proliferation,migration and invasion of A549 cells.TUG1 negatively regulated the expression of miR-142-5p,and miR-142-5p negatively regulated the expression of PD-L1,inhibition of miR-142-5p expression could reverse the inhibitory effects of silencing TUG1 on the proliferation,migration and invasion of A549 cells.Conclusion TUG1 is highly expressed in NSCLC tissues and cells,and silencing of TUG1 inhibits the proliferation,migration and invasion of A549 cells through the miR-142-5p/PD-L1 axis.