王 剑,陈清勇,张晓敏.转染microRNA-330-5p对肺癌细胞侵袭和迁移能力的影响[J].肿瘤学杂志,2014,20(8):648-653.
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
投稿时间:2014-04-30  
DOI:10.11735/j.issn.1671-170X.2014.08.B008
中文关键词:  miR-330-5p  哺乳动物雷帕霉素靶蛋白(mTOR)  肺肿瘤  侵袭  增殖
英文关键词:miR-330-5p  mammalian target of rapamycin(mTOR)  lung neoplasms  infiltration  proliferation
基金项目:南京军区“334”高层次科技人才培养工程;浙江省科技计划项目(2013C33209);杭州市医疗卫生科研项目(20130633B29)
作者单位
王 剑 浙江中医药大学第二临床医学院 
陈清勇 中国人民解放军第一一七医院 
张晓敏 浙江中医药大学第二临床医学院 
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中文摘要:
      摘 要:[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。
英文摘要:
      Abstract:[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 95C and 95D 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 95D and was significantly lower than that in 95C. 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 95D cells obviously decreased,the 24h 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 95C obviously increased,the 24h 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.
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