miR-181-5p下调人非小细胞肺癌细胞KLF6表达并抑制其增殖、迁移和侵袭

miR-181-5p down-regulates the expression of human non-small cell lung cancer cell KLF6 and inhibits its proliferation,migration and invasion

[目的]探讨miR-181-5p下调人非小细胞肺癌细胞KLF6表达并抑制其增殖、迁移和侵袭的作用。[方法]检测30例肺癌和癌旁组织中miR-181-5p和KLF6 mRNA水平,并分析miR-181-5p与NSCLC患者临床病理特征的相关性。用萤光素酶检测miR-181-5p对KLF6的调控作用,将miR-NC、miR-181-5p inhibitor和miR-181-5p mimic转染到A549细胞,采用RT-PCR法检测各组A549细胞中miR-181-5p和KLF6 mRNA表达,同时分别采用MTT法和Transwell法检测各组A549细胞增殖、迁移和侵袭情况。[结果]肺癌组织的miR-181-5p和KLF6 mRNA表达水平均低于癌旁组织(P<0.05);miR-181-5p的表达水平与年龄、性别和是否吸烟不相关(P>0.05);miR-181-5p的表达水平与肿瘤直径、是否转移、TNM分期相关(P<0.05);miR-181-5p与KLF6有潜在的结合位点;转染miR-181-5p mimic可明显增加KLF6-wt的荧光素酶活性,对KLF6-mut没有影响,而miR-181-5p inhibitor则表现出相反作用。给予miR-181-5p inhibitor干预后,细胞中miR-181-5p和KLF6 mRNA表达降低,细胞增殖率、迁移细胞数和侵袭细胞数增加(P<0.05);给予miR-181-5p mimic干预后,细胞中miR-181-5p和KLF6 mRNA表达增加,细胞增殖率、迁移细胞数和侵袭细胞数降低(P<0.05)。[结论]过表达miR-181-5p能抑制A549细胞增殖、迁移和侵袭,其机制可能与miR-181-5p通过靶向KLF6 mRNA的3'端非编码区,提高KLF6的翻译水平有关。

[Objective]To investigate the effect of miR-181-5p down-regulates the expression of human non-small cell lung cancer cell KLF6 and inhibits its proliferation,migration and invasion.[Method]The levels of miR-181-5p and KLF6 mRNA in 30 cases of lung cancer and adjacent tissues were detected,and the correlation between miR-181-5p and clinicopathological characteristics of NSCLC patients was analyzed.Luciferase was used to detect the regulation of miR-181-5p on KLF6.miR-NC,miR-181-5p inhibitor and miR-181-5p mimic were transfected into A549 cells,and RT-PCR method was used to detect the expression of miR-181-5p and KLF6 mRNA in A549 cells in each group.At the same time,MTT method and Transwell method were used to detect the proliferation,migration and invasion of A549 cells in each group.[Result]The expression levels of miR-181-5p and KLF6 mRNA in lung cancer tissues were lower than those in adjacent tissues(P<0.05).The expression levels of miR-181-5p were not related to age,gender and smoking status(P>0.05),and the expression level of miR-181-5p was related to tumor diameter,metastasis,TNM stage(P<0.05).miR-181-5p had potential binding sites with KLF6.miR-181-5p mimic transfection can significantly increase the luciferase activity of KLF6-wt,but had no effect on KLF6-mut,while miR-181-5p inhibitor shows the opposite effect.After the intervention of miR-181-5p inhibitor,the expression of miR-181-5p and KLF6 mRNA in the cell decreases,and the cell proliferation rate,the number of migrating cells and the number of invasive cells increased(P<0.05).After miR-181-5p mimic intervention,the expression of miR-181-5p and KLF6 mRNA in the cells increased,the cell proliferation rate,the number of migrating cells and the invasive cells decreased(P<0.05).[Conclusion]Overexpression of miR-181-5p can inhibit the proliferation,migration and invasion of A549 cells,and its mechanism may be related that miR-181-5p increases the translation level of KLF6 by targeting the 3'non-coding region of KLF6 mRNA.