miR-138-5p通过PDK1对人肺癌细胞系H23生物学行为的影响

Effect of mir-138-5p on biological behavior of human lung adenocarcinoma cell line H23 through PDK1

目的探讨微小RNA-138-5p(miR-138-5p)通过3-磷酸肌醇依赖性蛋白激酶1(PDK1)对人肺腺癌细胞系H23生物学行为的影响。方法以肺腺癌细胞系H23作为研究对象,分别转染miR-NC(对照组)或miR-138-5p(实验组);qRT-PCR与Western blot法检测miR-138-5p、PDK1 mRNA在转染后人肺腺癌细胞系H23中表达水平;双荧光素酶报告基因检测miR-138-5p及基因PDK1之间的靶向关系;MTT法、划痕实验、Transwell实验检测增殖、迁移与侵袭能力。结果qRT-PCR检测H23细胞转染后miR-138-5p mRNA表达水平升高,PDK1 mRNA与蛋白表达下调,荧光素酶报告基因实验提示miR-138-5p可与PDK1的3’-UTR直接结合,与PDK1存在靶向关系。miR-138-5p可抑制H23细胞增殖、迁移和侵袭能力。结论miR-138-5p可通过靶向干扰PDK1抑制人肺腺癌细胞系H23细胞的增殖、迁移与侵袭能力,提示miR-138-5p与PDK1可能是治疗肺癌的潜在靶点。

Objective To investigate the effect of microrna-138-5p(mir-138-5p)on the biological behavior of human lung cancer cell line H23 through inositol 3-phosphate dependent protein kinase 1(PDK1).Methods qRT-PCR and Western blot were used to detect the expression of mir-138-5p and PDK1 mRNA in human lung cancer cell line H23.Double luciferase reporter gene was used to detect the targeting relationship between mir-138-5p and PDK1.MTT assay,scratch test and Transwell test were used to detect the proliferation,migration and invasion ability.Results The expression of miR-138-5p mRNA in H23 cells was detected by qRT-PCR,PDK1 mRNA and protein expression were down regulated.Luciferase reporter gene experiment showed that miR-138-5p could directly bind to 3'-UTR of PDK1,and had a targeting relationship with PDK1.Conclusion MiR-138-5p can inhibit the proliferation,migration and invasion of human lung cancer cell line H23 by interfering with PDK1,indicating that mir-138-5p and PDK1 may be potential targets for the treatment of lung cancer.