miR-29a-3p和miR-144a-3p下调人非小细胞肺癌细胞COL11A1表达并抑制其增殖、迁移和侵袭

miR-29a-3p and miR-144a-3p down-regulate COL11A1 expression and inhibit proliferation,migration and invasion of human non-small cell lung cancer cells

目的探讨XI型胶原α1(collagen type XI alpha 1,COL11A1)、miR-29a-3p和miR-144a-3p在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的作用和它们之间的内在联系。方法用RT-qPCR法检测NSCLC组织和细胞中COL11A1 mRNA和与COL11A1具有可能靶向关系的miRNAs的表达水平。COL11A1 siRNA、miR-29a-3p mimic或miR-144a-3p mimic转入H520细胞后,CCK-8法检测细胞活性,EdU法检测细胞增殖,Transwells法检测细胞迁移和侵袭,Western blot法检测COL11A1、Ki67和MMP-2的蛋白表达水平。荧光素酶报告基因分析检测COL11A1是否是miR-29a-3p和miR-144a-3p在NSCLC细胞中的靶点。结果在NSCLC肿瘤组织和细胞中,CoL11A1的表达水平均明显上调(P<0.01),miR-29a-3p和miR-144a-3p的表达水平均明显下调(P<0.01)。敲低COL11A1或过表达miR-29a-3p和miR-144a-3p均能抑制H520细胞增殖、迁移和侵袭(P<0.05或P <0.01),且过表达miR-29a-3p和miR-144a-3p均能抑制COL11A1表达。荧光素酶报告基因实验证实miR-29a-3p和miR-144a-3p均可直接靶向COL11A1。结论过表达miR-29a-3p和miR-144a-3p可通过直接下调COL11A1表达来抑制NSCLC细胞的增殖、迁移和侵袭。

Aim To explore the role and the internal relations of collagen type XI alpha 1(COL11A1),miR-29a-3p and miR-144a-3p in non-small cell lung cancer(NSCLC).Methods The expression levels of COL11A1 mRNA and miRNAs with a possible targeting relationship with COL11A1 in NSCLC tissues and cells were detected by RT-qPCR.After transfected with COL11A1 siRNA,miR-29a-3p mimic or miR-144a-3p mimic into H520 cells,cell viability was detected by CCK-8 assay,cell proliferation was detected by EdU assay,cell migration and invasion were detected by Transwells assay,and the expression levels of COL11A1,Ki67 and MMP-2 were detected by Western blot.Luciferase reporter gene analysis was used to determine whether COL11A1 was the target of miR-29a-3p and miR-144a-3p in NSCLC cells.Results COL11A1 expression levels were significantly up-regulated in NSCLC tissues and cell lines(P<0.01),and the miR-29a-3p and miR-144a-3p levels were dramatically down-regulated(P<0.01).Knockdown of COL11A1 or over-expression of miR-29a-3p and miR-144a-3p inhibited the proliferation,migration and migration of H520 cells(P<0.05 or P<0.01),and over-expression of miR-29a-3p and miR-144a-3p inhibited COL11A1 expression(P<0.01).miR-29a-3p and miR-144a-3p could directly target COL11A1 by using luciferase reporter assay.Conclusions Over-expression of miR-29a-3p and miR-144a-3p inhibits cell proliferation migration and invasion of NSCLC cells by directly down-regulating COL11A1 expression.