lncRNA HOXC-AS3吸附miR-216a-5p促进非小细胞肺癌细胞增殖和迁移的机制研究

Mechanism research of lncRNA HOXC-AS3 inhibiting cell proliferation,migration and invasion of non-small cell lung cancer cells by targeting miR-216a-5p

目的:探讨lncRNA HOXC-AS3对非小细胞肺癌(NSCLC)细胞增殖、迁移和侵袭的影响及分子机制。方法:HOXC-AS3小干扰RNA(si-HOXC-AS3)、HOXC-AS3过表达载体(pcDNA-HOXC-AS3)或miR-216a-5p抑制剂(anti-miR-216a-5p)Western blot检测细胞增殖抗原Ki-67、细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和上皮型钙黏蛋白(E-cadherin)表达,双荧光素酶报告系统验证lncRNA H1299、A549和GLC-12中lncRNA HOXC-AS3水平上升,miR-216a-5p水平降低(P<0.05);沉默lncRNA HOXC-AS3可抑制A549高E-cadherin蛋白表达,过表达lncRNA HOXC-AS3可促进A549细胞增殖、迁移、侵袭和EMT;lncRNA HOXC-AS3靶向吸附cRNA HOXC-AS3通过吸附miR-216a-5p促进NSCLC A549细胞增殖、迁移、侵袭和EMT,是肺癌的潜在分子靶点。

Objective:To investigate effects of lncRNA HOXC-AS3 on proliferation,migration and invasion of non-small cell lung cancer(NSCLC)cells and molecular mechanism.Methods:Levels of lncRNA HOXC-AS3 and miR-216 a-5 p in lung cancer tissues and H1299,A549 and GLC-12 cells were detected by qRT-PCR.A549 cells were transfected with si-HOXC-AS3,pcDNAHOXC-AS3 or anti-miR-216 a-5 p to silence,overexpress lncRNAHOXC-AS3 or interfere miR-216 a-5 p.Cell proliferation of A549 cells was detected by CCK-8 assay.Cell migration and invasion of A549 cells were detected by Transwell assay and scratch test.Protein expressions of Ki-67,Cyclin D1,MMP-2,MMP-9,N-cadherin,Vimentin and E-cadherin were determined by Western blot.Dual-luciferase reporter assay system was performed to verify targeting relationship between lncRNA HOXC-AS3 and miR-216 a-5 p.Results:Compared with adjacent tissues(45 cases)and human normal lung epithelial cells BEAS-2 B,levels of lncRNA HOXC-AS3 in lung cancer tissues and lung cancer H1299,A549 and GLC-12 cells were increased,while levels of miR-216 a-5 p were decreased(P<0.05).Silencing lncRNA HOXC-AS3 inhibited proliferation,migration,invasion and epithelial-mesenchymal transition(EMT)of A549 cells,reduced protein expressions of Ki-67,Cyclin D1,MMP-2,MMP-9,N-cadherin and Vimentin,and increased expression of E-cadherin.Over-expression of lncRNA HOXC-AS3 promoted proliferation,migration,invasion and EMT of A549 cells.lncRNA HOXC-AS3 sponged and regulated expression of miR-216 a-5 p.Interference with miR-216 a-5 p partially reversed inhibitory effect of lncRNA HOXC-AS3 silenced on proliferation,migration,invasion and EMT of A549 cells.Conclusion:lncRNA HOXC-AS3 promoted proliferation,migration and invasion of A549 cells by targeting miR-216 a-5 p,which is a potential molecular target for NSCLC.