miR-519d-3p靶向TROAP调节Wnt/β-Catenin信号通路对非小细胞肺癌增殖、凋亡、迁移和侵袭的影响

Effects of miR-519d-3p targeting TROAP to regulate Wnt/β-Catenin signaling pathway on proliferation,apoptosis,migration,and invasion in non-small cell lung cancer

目的:探究微小RNA-519d-3p(miR-519d-3p)是否靶向肌钙蛋白相关蛋白(TROAP)调节Wnt/β-Catenin信号通路影响非小细胞肺癌(NSCLC)细胞增殖、凋亡、迁移和侵袭。方法:RT-qPCR评估miR-519d-3p和TROAP mRNA表达;双荧光素酶分析miR-519d-3p与TROAP之间的靶向关系;Western blotting检测TROAP、β-Catenin、Axin2、C-myc的蛋白表达;集落形成实验、流式细胞术和Transwell法检测A549细胞增殖、凋亡、迁移和侵袭;BABL/c裸鼠建立移植瘤模型,检测miR-519d-3p对体内肿瘤生长的影响。结果:miR-519d-3p与TROAP存在靶向关系(P<0.05);在体外,miR-519d-3p过表达可抑制A549细胞增殖、迁移和侵袭,并促进细胞凋亡。此外,miR-519d-3p过表达可下调TROAP表达并降低β-Catenin、Axin2、C-myc蛋白水平(P<0.05);TROAP过表达对A549细胞的作用与之相反(P<0.05)。同时,过表达TROAP可部分逆转miR-519d-3p过表达对A549细胞恶性行为的影响(P<0.05)。在体内,miR-519d-3p过表达可减小肿瘤体积和肿瘤重量,且显著下调Ki67和N-cadherin表达(P<0.05)。结论:miR-519d-3p靶向TROAP抑制Wnt/β-Catenin信号通路,抑制NSCLC恶性生物学行为。

Objective:To investigate whether microRNA-519d-3p(miR-519d-3p)targets trophinin associated protein(TROAP)to regulate Wnt/β-Catenin signaling pathway to affect proliferation,apoptosis,migration,and invasion of non-small cell lung cancer(NSCLC)cells.Methods:Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to assess miR-519d-3p and TROAP mRNA expression;dual luciferase was used to analyze the targeting relationship between miR-519d-3p and TROAP;Western blotting was used to detect the protein expression of TROAP,β-Catenin,Axin2,C-myc;colony formation assay,flow cytometry and Transwell assays was used to detect proliferation,apoptosis,migration,and invasion of A549 cells;transplantation tumor model established using BABL/c nude mice to detect the effect of miR-519d-3p on tumor growth in vivo.Results:miR-519d-3p had a targeting relationship with TROAP(P<0.05).In vitro,miR-519d-3p overexpression inhibited A549 cell proliferation,migration and invasion and promoted apoptosis;in addition,miR-519d-3p overexpression downregulated TROAP expression and decreasedβ-Catenin,Axin2,and C-myc protein levels(P<0.05);TROAP overexpression had the opposite effect on A549 cells(P<0.05).Meanwhile,overexpression of TROAP partially reversed the effect of miR-519d-3p overexpression on the malignant behavior of A549 cells(P<0.05).In vivo,miR-519d-3p overexpression reduced tumor volume and tumor weight,and significantly downregulated Ki67 and N-cadherin expression(P<0.05).Conclusion:miR-519d-3p targets TROAP to inhibit Wnt/β-catenin signaling pathway and suppress malignant biological behaviors of NSCLC.