LncRNA TMPO-AS1通过调节miR-204-3p影响肺癌细胞的增殖、凋亡和侵袭  

LncRNA TMPO-AS1 affects proliferation,apoptosis and invasion of lung cancer cells by modulating miR-204-3p

作　　者：邓俊亮 华美香 陈键腾 DENG Jun-Liang;HUA Mei-Xiang;CHEN Jian-Teng(Department of Respiratory Medicine,Xinhui Hospital,Southern Medical University,Jiangmen 529100,China)

机构地区：[1]南方医科大学附属新会医院呼吸内科,江门529100 [2]南方医科大学附属新会医院神经内科,江门529100

出　　处：《中国免疫学杂志》2021年第3期319-324,329,共7页Chinese Journal of Immunology

摘　　要：目的:探究长链非编码RNA(lncRNA)TMPO-AS1通过调节miR-204-3p的表达对肺癌细胞增殖、凋亡和侵袭的影响。方法:实时荧光定量PCR(qPCR)检测人肺癌细胞系A549、H1975、H1299和H1650和人正常支气管上皮细胞系HBE中TMPO-AS1的表达水平。采用脂质体转染技术沉默A549细胞中TMPO-AS1,qPCR检测TMPO-AS1和miR-204-3p的表达情况,荧光素酶报告实验检测TMPO-AS1和miR-204-3p的靶向关系,噻唑蓝比色法(MTT)、流式细胞术和Transwell实验分别检测沉默TMPO-AS1对A549细胞增殖、凋亡和侵袭能力的影响,Western blot检测A549细胞中PCNA、Ki-67、Cleaved Caspase-3、Cleaved Caspase-9、MMP-2和MMP-9的表达。结果:人肺癌细胞系中TMPO-AS1的表达水平明显高于人正常支气管上皮细胞系(P<0.05),且肺癌A549细胞中TMPO-AS1的基础表达量最高,选择A549细胞用于后续实验。沉默TMPO-AS1可明显促进miR-204-3p的表达(P<0.05),荧光素酶报告实验显示,TMPO-AS1能够降低miR-204-3p野生型细胞的相对荧光素酶活性(P<0.05)。沉默TMPO-AS1能够明显抑制A549细胞增殖和侵袭能力(P<0.05),降低增殖相关蛋白PCNA、Ki-67及侵袭相关蛋白MMP-2、MMP-9的表达(P<0.05);此外,沉默TMPO-AS1可促进A549细胞凋亡及Cleaved Caspase-3和Cleaved Caspase-9的表达(P<0.05)。结论:沉默TMPO-AS1能够通过靶向促进miR-204-3p的表达抑制肺癌细胞增殖和侵袭,并诱导细胞凋亡。Objective:To investigate the effects of long-chain non-coding RNA(lncRNA)TMPO-AS1 on proliferation,apoptosis and invasion of lung cancer cells by regulating the expression of miR-204-3p.Methods:Real-time quantitative PCR(qPCR)was used to detect the expression levels of TMPO-AS1 in human lung cancer cell lines A549,H1975,H1299 and H1650 and human normal bronchial epithelial cell line HBE.The liposome transfection technique was used to silence of TMPO-AS1 in A549 cells.The expression of TMPO-AS1 and miR-204-3p was detected by qPCR.The luciferase reporter assay was used to detect the targeting relationship between TMPO-AS1 and miR-204-3p.Thiazole blue colorimetric assay(MTT),flow cytometry and Transwell assay were used to detect the effect of silencing TMPO-AS1 on proliferation,apoptosis and invasion of A549 cells.Western blot was used to detect expressions of PCNA,Ki-67,Cleaved Caspase-3,Cleaved Caspase-9,MMP-2 and MMP-9 in A549 cells.Results:The expression level of TMPO-AS1 in human lung cancer cell line was significantly higher than that in human normal bronchial epithelial cell line(P<0.05),and the expression of TMPO-AS1 was the highest in lung cancer A549 cells.A549 cells were selected for subsequent experiments.Silencing TMPO-AS1 significantly promoted the expression of miR-204-3p(P<0.05).Luciferase reporter assay showed that TMPO-AS1 could decrease the relative luciferase activity of miR-204-3p wild-type cells(P<0.05).Silencing TMPO-AS1 significantly inhibited the proliferation and invasion of A549 cells(P<0.05),and decreased the expression of proliferation-related proteins PCNA,Ki-67 and invasion-related proteins MMP-2 and MMP-9(P<0.05).TMPO-AS1 promoted apoptosis of A549 cells and expression of Cleaved Caspase-3 and Cleaved Caspase-9(P<0.05).Conclusion:Silencing TMPO-AS1 can inhibit the proliferation and invasion of lung cancer cells and induce apoptosis by targeting the expression of miR-204-3p.

关 键 词：LncRNA TMPO-AS1 miR-204-3p 肺癌细胞 增殖 凋亡 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]