LncRNA PSMA3-AS1调控miR-27b-3p对非小细胞肺癌细胞增殖、凋亡、迁移的影响  

LncRNA PSMA3-AS1 regulates cell proliferation,apoptosis,and migration of non-small cell lung cancer cells by mediating miR-27b-3p

作　　者：魏童[1] 王学成 赵亮[1] WEI Tong;WANG Xuecheng;ZHAO Liang(Department of Cardiothoracic Surgery,the Fifth Affiliated Hospital of Xinjiang Medical University,Xinjiang,Urumqi 830011,China)

机构地区：[1]新疆医科大学第五附属医院心胸外科,乌鲁木齐市830011

出　　处：《河北医药》2023年第12期1765-1769,共5页Hebei Medical Journal

基　　金：新疆维吾尔自治区自然科学基金项目(编号:2022D01C328)。

摘　　要：目的探究长链非编码RNA(lncRNA)人蛋白酶体α亚基3型反义RNA1(PSMA3-AS1)通过调控微小RNA-27b-3p(miR-27b-3p)对非小细胞肺癌细胞增殖、凋亡、迁移的影响。方法选取20例非小细胞肺癌标本和20例健康志愿者肺部上皮样本。购买人体正常的肺部上皮细胞H1299、H358、A549及NC-LH2073.采用实时荧光RT-PCR检测LncRNA PSMA3-AS1、miR-27b-3p表达,构建抑制LncRNA PSMA3-AS1表达的A549细胞,采用MTT法、流式细胞仪、Transwell检测细胞增殖、凋亡、迁移情况,Western Blot法检测细胞周期蛋白-D1(CyclinD1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达变化。结果与人体正常肺上皮组织相比,非小细胞肺癌组织中LncRNA PSMA3-AS1表达升高,miR-27b-3p表达下降(P<0.05)。与BEAS-2B组相比,H1299组、H358组、NC-LH2073组、A549组中LncRNA PSMA3-AS1水平上升,miR-27b-3p水平降低(P<0.05)。与si-NC组相比,si-PSMA3-AS1组lncRNA PSMA3-AS1水平表达降低(P<0.05),与si-NC组相比,si-HIF-1a组OD值、CyclinD1、MMP-2、MMP-9、迁移细胞数下降,凋亡率升高(P<0.05)。与miR-NC组相比,miR-27b-3p组miR-27b-3p表达水平降低(P<0.05),与miR-NC组相比,miR-27b-3p组OD值、CyclinD1、MMP-2、MMP-9、迁移细胞数降低,凋亡率上升(P<0.05)。双荧光素酶报告试验显示,LncRNA PSMA3-AS1靶向miR-27b-3p表达,且经转染后,通过RT-qPCR检测显示miR-NC组及miR-27b-3p组中的miR-27b-3p表达差异有统计学意义(P<0.05)。双荧光素酶报告发现同时转染miR-27b-3p、WT-PSMA3-AS1后,荧光素酶活性下降(P<0.05)。与si-NC组相比,抑制lncRNA-PSMA3-AS1表达可使PC3细胞中miR-27b-3p表达降低(P<0.05),与si-PSMA3-AS1+anti-miR-NC组相比,si-PSMA3-AS1+anti-miRNA-27b-3p组OD值、CyclinD1、MMP2、MMP9、迁移细胞数上升,凋亡率降低(P<0.05)。结论干扰LncRNA PSMA3-AS1水平表达,可对miR-27b-3p进行有效调控,从而抑制非小细胞肺癌细胞的增殖与迁移,加速其凋亡,阻碍非小细胞肺癌的恶性发展。Objective To explore the regulatory effect of long non-coding RNA(lncRNA)human proteasomalαsubunit 3 antisense RNA1(PSMA3 AS1)on cell proliferation,apoptosis and migration of non-small cell lung cancer(NSCLC)cells by mediating miR-27b-3p.Methods A total of 20 NSCLC specimens and 20 lung epithelium specimens of healthy volunteers were collected.Expression levels of lncRNA PSMA3-AS1 and miR-27b-3p in them,NSCLC cell lines H1299,H358,A549 and NCI-H2073,and the A549 cells that inhibited the expression of LncRNA PSMA3-AS1 were detected by quantitative real-time PCR(qRT-PCR).The cell proliferation,apoptosis and migration were detected by MTT assay,flow cytometry and Transwell assay,respectively.Protein expressions of Cyclin D1,matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9)were detected by Western Blot.Results Compared with those of human normal lung epithelial tissues,lncRNA PSMA3-AS1 was upregulated,and miR-27b-3p was downregulated in NSCLC tissues(P<0.05).Compared with those of BEAS-2B cells,lncRNA PSMA3-AS1 was upregulated,and miR-27b-3p was downregulated in NSCLC cell lines H1299,H358 and NCI-H2073(P<0.05).Transfection of si-PSMA3-AS1 significantly downregulated lncRNA PSMA3-AS1,OD value,Cyclin D1,MMP-2 and MMP-9,reduced migratory cell number and increased apoptotic rate(P<0.05).Transfection of miR-27b-3p inhibitor significantly downregulated miR-27b-3p,OD value,Cyclin D1,MMP-2 and MMP-9,reduced migratory cell number and increased apoptotic rate(P<0.05).Dual-luciferase reporter assay revealed that lncRNA PSMA3-AS1 targeted miR-27b-3p.Co-transfection of miR-27b-3p inhibitor and WT-PSMA3-AS1 significant reduced luciferase activity(P<0.05).Transfection of si-PSMA3-AS1 significantly downregulated miR-27b-3p(P<0.05).Co-transfection of si-PSMA3-AS1 and anti-miR-27b-3p significantly increased the proliferative rate and migratory cell number,upregulated OD value,Cyclin D1,MMP-2 and MMP-9,and reduced apoptotic rate compared with those co-transfected with si-PSMA3-AS1 and anti-miR-NC(P<0.05).Conclusi

关 键 词：非小细胞肺癌 LncRNA PSMA3-AS1 微小RNA-27b-3p 增殖 凋亡 迁移 

分 类 号：R734.2[医药卫生—肿瘤]