lncRNA-miR210HG在非小细胞肺癌发生中的作用机制

The mechanism of lncRNA-miR210HG in the occurrence of NSCLC

目的:探讨lncRNA-miR210HG在非小细胞肺癌(NSCLC)发生中的作用机制。方法:收集2017年01月至2020年01月我院收治确诊的54例NSCLC患者的癌组织与癌旁组织,采用生物信息学在线程序预测lncRNA-miR210HG的潜在靶标miRNA,双荧光素酶实验检测NCI-H1650 NSCLC细胞中lncRNA-miR210HG和miRNA210的相互作用,qRT-PCR检测肺癌组织中lncRNA-miR210HG、miRNA210、HIF-1α、VEGF的mRNA表达水平,NCI-H1650 NSCLC细胞分别转染miRNA210类似物及抑制物后,采用qRT-PCR检测细胞水平lncRNA-miR210HG、HIF-1α与VEGF的mRNA表达。结果:lncRNA-miR210HG与miRNA210间存在互补结合序列,野生型lncRNA-miR210HG和miRNA210的荧光素酶活性低于对照组(P<0.01),且lncRNA-miR210HG与miRNA210的靶向结合是通过其3'-UTR区。NSCLC组织中lncRNA-miR210HG、miRNA210、HIF-1α(P<0.001)及VEGF(P<0.01)的mRNA表达水平均较癌旁组织中高,差异有统计学意义。NSCLC组织中lncRNA-miR210HG表达与miRNA210表达呈负相关。细胞实验发现,miRNA210过表达后,HIF-1α、VEGF、lncRNA-miR210HG的mRNA表达水平均下降,其中lncRNA-miR210HG、HIF-1α与对照组相比差异有统计学意义(lncRNA-miR210HG:P<0.05;HIF-1α:P<0.01),miRNA210被抑制后,HIF-1α、VEGF、lncRNA-miR210HG的mRNA表达水平均较对照组升高,且差异均有统计学意义(lncRNA-miR210HG:P<0.01;HIF-1α:P<0.05;VEGF:P<0.01)。结论:lncRNA-miR210HG 3'-UTR区通过与miRNA210的靶向结合,从而调控下游HIF-1α、VEGF的mRNA表达,可能参与了NSCLC的发生、发展。

Objective:To explore the mechanism of lncRNA-miR210HG in the occurrence of non-small cell lung cancer.Methods:The cancer and adjacent tissues of 54 cases of non-small cell lung cancer patients diagnosed in our hospital from January 2017 to January 2020 were collected.Using bio-informatics online program to predict the potential target miRNA of lncRNA-miR210HG,and the interaction of lncRNA-miR210HG and miRNA210 in lung cancer cells was detected by dual luciferase assay.qRT-PCR was used to detect the mRNA expression levels of lncRNA-miR210HG,miRNA210,HIF-1αand VEGF in lung cancer tissues.After NCI-H1650 NSCLC cells were transfected with miRNA210 analogs and inhibitors,qRT-PCR was used to detect the mRNA expression levels oflncRNA-miR210HG,HIF-1αand VEGF at the cellular level.Results:There was a complementary binding sequence between lncRNA-miR210HG and miRNA210.The luciferase activity in lncRNA-miR210HG WT andmiRNA210 was lower than the control group(P<0.01),and the targetedbinding of lncRNA-miR210HG andmiRNA210 was through its 3'-UTR region.The mRNA expression levelsof lncRNA-miR210HG,miRNA210,HIF-1α(P<0.001)and VEGF(P<0.01)in non-small cell lung cancer tissues were higher than those in adjacent tissues,and the difference was statistically significant.The expression of lncRNA-miR210HG in non-small cell lung cancer tissues wasnegatively correlated with the expression of miRNA210.Cell experiments found that,aftermiRNA210 was overexpressed,the mRNA expression levels of HIF-1α,VEGF and lncRNA-miR210HG decreased.Among them,lncRNA-miR210HG and HIF-1αwere significantly different from the control group(lncRNA-miR210HG:P<0.05,HIF-1α:P<0.01),while after miRNA210 was inhibited,the mRNA expression levels ofHIF-1α,VEGF and lncRNA-miR210HG were higher than those of the control group,and the differences were statistically significant(lncRNA-miR210HG:P<0.01,HIF-1α:P<0.05,VEGF:P<0.01).Conclusion:The 3'-UTR region of lncRNA-miR210HG may bind to miRNA210,thereby regulating downstream HIF-1αand VEGF mRNA expression,and participating in the occurrence and development of NSCLC.