LINC00691负调控miR-512-5p对非小细胞肺癌细胞增殖、凋亡、迁移和侵袭的影响

Effects of the negative regulation of miR-512-5p by LINC00691 on the proliferation, apoptosis, migration and invasion of NSCLC cells

摘要:

目的 探讨LINC00691对非小细胞肺癌（NSCLC）细胞恶性生物学行为的影响。方法 RT-qPCR检测30例NSCLC患者癌组织和对应癌旁组织中LINC00691和miR-512-5p的表达。双荧光素酶报告基因实验验证LINC00691和miR-512-5p的靶向关系。将A549细胞分为si-NC组、si-LINC00691组、miR-NC组、miR-512-5p组、si-LINC00691+anti-miR-NC组和si-LINC00691+anti-miR- 512-5p组，MTT和克隆形成实验检测细胞增殖情况，流式细胞术、Transwell检测细胞凋亡、迁移和侵袭，Western blotting检测细胞中Ki67、Cleaved-caspase3、E-cadherin和N-cadherin蛋白表达。结果 LINC00691在NSCLC癌组织中表达水平上升（P<0.05），而miR-512-5p表达水平下降（P<0.05）。LINC00691在A549细胞中负调控miR-512-5p。沉默LINC00691表达或过表达miR-512-5p可降低A549细胞的存活率、克隆形成数、迁移和侵袭数以及Ki67、N-cadherin蛋白表达水平，提高其凋亡率，上调Cleaved-caspase3、E-cadherin蛋白表达水平（P<0.05）。沉默miR-512-5p表达逆转了沉默LINC00691表达对A549细胞增殖、迁移和侵袭的抑制作用及凋亡促进作用。结论 沉默LINC00691可能通过靶向上调miR-512-5p表达来抑制NSCLC细胞的增殖、迁移和侵袭，并促进其凋亡。

Abstract:

Objective To investigate the effects of LINC00691 on the malignant biological behavior of non-small cell lung cancer (NSCLC) cells.Methods RT-qPCR was used to detect the expressions of LINC00691 and miR-512-5p in 30 cases of NSCLC tissues and corresponding adjacent tissues. The dual luciferase reporter gene assay was used to verify the targeting relationship between LINC00691 and miR-512-5p. NSCLC A549 cells were cultured in vitro, and were designed into si-NC group, si-LINC00691 group, miR-NC group, miR-512-5p group, si-LINC00691+anti-miR-NC group and si-LINC00691+anti-miR-512-5p group. MTT and clone formation experiment were applied to detect the cell proliferation, and flow cytometry to detect cell apoptosis, and Transwell to detect cell migration and invasion. Western blotting was used to detect the expressions of Ki67, cleaved caspase-3, E-cadherin and N-cadherin proteins.Results The expression of LINC00691 increased in NSCLC cancer tissues (P<0.05), while the expression of miR-512-5p decreased (P<0.05). LINC00691 negatively regulated the miR-512-5p in A549 cells. Silenced LINC00691 or over-expressed miR-512-5p promoted the apoptosis rate and restrained the viability, colony formation, migration and invasion of A549 cells, down-regulated the expressions of Ki67 and N-cadherin proteins (P<0.05), but up-regulated the expressions of cleaved-caspase3 and E-cadherin proteins (P<0.05). Silenced miR-512-5p reversed the inhibitory effects of silenced LINC00691 on the proliferation, migration and invasion of A549 cells, as well as their apoptosis-promoting effects.Conclusion Silenced LINC00691 could inhibit the proliferation, migration and invasion, and promote the apoptosis of NSCLC cells by targeting up-regulation of miR-512-5p.