转染microRNA-330-5p对肺癌细胞侵袭和迁移能力的影响

Impact of Transfection of MiR-330-5p on the Invasion and Metastasis of Lung Cancer Cells

[目的]探讨miR-330-5p表达对肺癌细胞增殖、侵袭和迁移能力等生物学行为的影响,揭示miR-330-5p的作用机制。[方法]分析非小细胞肺癌(NSCLC)细胞系95C和95D的miRNA表达谱芯片中miR-330-5p的表达情况并用靶标软件预测其靶基因;转染miR-330-5p类似物(mimics)至95D、转染miR-330-5p抑制剂(inhibitor)至95C中,应用CCK-8法、transwell小室法检测miR-330-5p对肺癌细胞增殖、侵袭迁移等生物学行为的影响;应用Western blot法检测miR-330-5p表达对哺乳动物雷帕霉素靶蛋白(mTOR)表达水平的影响。[结果]miRNA表达谱芯片显示miR-330-5p在95D中表达明显低于95C;靶标预测软件预测mTOR mRNA可能是miR-330-5p的靶基因;转染miR-330-5p mimics后95D细胞增殖能力显著降低,其24h侵袭穿膜细胞数明显低于对照组,mTOR蛋白表达明显下调(P<0.01);而转染miR-330-5p inhibitor后95C细胞的增殖能力增强,其24h侵袭穿膜细胞数较对照组明显增加,mTOR蛋白表达上调(P<0.01)。[结论]提高miR-330-5p表达能够明显抑制肺癌细胞的增殖、侵袭与迁移能力,mTOR mRNA可能是其重要的靶基因。

[Purpose] To investigate the effect of miR-330-5p on the proliferation,invasion and metastasis ability of lung cancer cells and to reveal the possible mechanisms of miR-330-5p. [Methods] The expression level of miR-330-5p in non-small cell lung cancer（NSCLC） cell lines 95 C and 95 D was detected by miRNA expression profile chip technology,and the target genes were predicted by target genes prediction software. MiR-330-5p analogue（mimics） was transfected to high metastatic lung cancer cells（95D）,while miR-330-5p（inhibitor） was transfected to low metastatic lung cancer cells（95C）.The proliferation,invasion and metastasis ability of lung cancer cells was detected by CCK-8 and Transwell Chambers method,and the expression level of mTOR protein was determined by Western blot method. [Results] The results of miRNA expression profile chip showed that the expression level of miR-330-5p down-regulated in 95 D and was significantly lower than that in 95 C. mTOR mRNA may be a target gene of miR-330-5p predicting by target genes prediction software. After transfected with miR-330-5p mimics,the proliferation ability of 95 D cells obviously decreased,the 24 h number of cells with infiltration of membrane was significantly lower than that of the controls（P〈0.01）,and the expression of mTOR protein down-regulated（P〈0.01）.While after transfected with miR-330-5p inhibitor,the proliferation ability of 95 C obviously increased,the 24 h number of cells with infiltration of membrane was significantly higher than that of the controls（P〈0.01）,and the expression of mTOR protein up-regulated（P〈0.01）.[Conclusion] Increasing the miR-330-5p expression level might significantly inhibit the proliferation,invasion and metastasis ability of lung cancer cells,and mTOR mRNA may be a target gene of miR-330-5p.