LINC00710靶向miR-367/PTEN轴抑制肺癌A549细胞的增殖、迁移和侵袭

LINC00710 targets miR-367/PTEN axis to inhibit the proliferation,migration,and invasion of lung cancer A549 cells

目的:探讨LINC00710对肺癌细胞A549增殖、迁移和侵袭的影响及其分子机制。方法:收集2017年12月至2018年12月本院收治的33例肺癌患者的癌组织及癌旁组织,RT-qPCR检测肺癌组织、癌旁组织中LINC00710和miR-367表达水平;双荧光素酶报告基因实验确定LINC00710对miR-367、miR-367对PTEN靶向调控作用。体外培养A549细胞,并将LINC00710过表达载体、miR-367模拟物(mimic)分别转染A549细胞,分为对照组、pcDNA3.1组、pcDNA3.1-LINC00710组、miR-367 mimic组、miR-367 mimic+pcDNA3.1-LINC00710组。CCK-8和克隆形成实验检测细胞增殖,Transwell实验检测细胞迁移和侵袭;Western blotting检测PTEN蛋白表达。结果:与癌旁组织相比,肺癌组织中LINC00710表达水平降低,miR-367表达水平升高(P<0.05)。LINC00710靶向负调控miR-367表达,miR-367靶向负调控PTEN表达。与对照组比较,pcDNA3.1-LINC00710组A549细胞增殖活性、克隆形成数、迁移及侵袭细胞数、miR-367、Ki67和N-cadherin表达水平降低,PTEN和E-cadherin表达升高(均P<0.05),miR-367 mimic组A549细胞增殖活性、克隆形成数、迁移和侵袭细胞数、miR-367、Ki67和N-cadherin表达水平升高,PTEN和E-cadherin表达降低(均P<0.05);与pcDNA3.1-LINC00710组比较,miR-367 mimic+pcDNA3.1-LINC00710组A549细胞增殖活性、克隆形成数、迁移和侵袭细胞数、miR-367、Ki67和N-cadherin表达水平升高,PTEN和E-cadherin表达降低(均P<0.05)。结论:LINC00710靶向miR-367/PTEN轴能够抑制肺癌A549细胞的增殖、迁移和侵袭。

Objective:To explore the effect of LINC00710 on the proliferation,migration,and invasion of lung cancer A549 cells and its molecular mechanism.Methods:We collected the cancerous and paracancerous tissue samples of 33 lung cancer patients admitted to our hospital from December 2017 to December 2018.The expression levels of LINC00710 and miR-367 in lung cancer tissue and paracancerous tissue were detected by real-time reverse transcription quantitative PCR.A dual luciferase reporter gene experiment was applied to confirm the targeted relationship between LINC00710 and miR-367,miR-367,and PTEN.A549 cells were cultured in vitro,and the LINC00710 overexpression vector and miR-367 mimic were transfected into A549 cells,respectively.The cells were divided into four groups,including a control group,pcDNA3.1 group,pcDNA3.1-LINC00710 group,miR-367 mimic group,and miR-367 mimic+pcDNA3.1-LINC00710 group.Both cell count kit(CCK-8)and clone formation assays were used to detect cell proliferation.Transwell assay was used to detect cell migration and invasion.Western blotting was applied to detect the protein expression level of PTEN.Results:Compared with adjacent tissue,the expression level of LINC00710 in lung cancer tissue was significantly reduced,while the expression level of miR-367 was significantly increased(P<0.05).LINC00710 targeted and negatively regulated miR-367 expression,miR-367 targeted and negatively regulated PTEN expression.Compared with the control group,the proliferation activity,colony formation number,migration and invasion cell number,miR-367 level,Ki67,and N-cadherin expression of A549 cells in the pcDNA3.1-LINC00710 group were decreased,while the PTEN and E-cadherin expression were increased(all P<0.05).When compared with the control group,A549 cell proliferation activity,colony formation number,migration and invasion cell number,miR-367 level,Ki67,and N-cadherin expression were increased in the miR-367 mimic group,but the PTEN and E-cadherin expression was decreased(all P<0.05).Compared with the pcDNA3.1-LINC00710 group,A549 cells in the miR-367 mimic+pcDNA3.1-LINC00710 group had increased proliferation activity,colony formation number,migration and invasion cell number,miR-367 level,Ki67,and N-cadherin expression;however,the PTEN and E-cadherin expression were decreased(all P<0.05).Conclusions:LINC00710 could inhibit the proliferation,migration,and invasion of lung cancer A549 cells by targeting the miR-367/PTEN axis.