miR-196a调控p27^kip1促进非小细胞肺癌细胞增殖的机制研究  

Up-regulated miR-196a promotes non-small cell lung cancer cells proliferation by regulating p27^(kip1) expression

作　　者：刘向华[1] 林梦洁[2] 尹凌帝 孙倩[2] 孙明[1] 德伟[1] 刘志军[1] 

机构地区：[1]南京医科大学生物化学与分子生物学系,南京210029 [2]南京医科大学第一临床医学院,210029

出　　处：《临床肿瘤学杂志》2013年第11期961-965,共5页Chinese Clinical Oncology

基　　金：国家自然科学基金资助项目(81070620)

摘　　要：目的探讨miR-196a在非小细胞肺癌(NSCLC)组织及细胞系中的表达,以及抑制或过表达miR-196a对NSCLC细胞增殖能力的影响及其作用的靶基因。方法实时定量PCR(QPCR)技术检测NSCLC组织及细胞系中miR-196a的表达水平,通过转染miR-196a inhibitors及miR-196a mimics抑制或上调miR-196a的表达水平,并通过QPCR检测转染效率。用MTT和克隆形成实验检测上调或下调miR-196a对SPC-A1或A549细胞增殖能力的影响;生物信息学及Western blotting方法分析验证miR-196a对靶基因p27kip1的调控作用。结果与正常肺组织及细胞相比,在NSCLC组织和细胞中miR-196a的表达出现显著上调,SPC-A1细胞和A549细胞中转染miR-196a inhibitors或miR-196a mimics能显著抑制或上调miR-196a的表达;抑制miR-196a的表达能降低SPC-A1细胞增殖能力,而上调miR-196a的表达则能促进A549细胞增殖。miR-196a能够负性调控p27kip1的表达。结论 NSCLC组织及细胞中miR-196a的表达上调能够抑制p27kip1的表达,并显著促进NSCLC细胞增殖,影响NSCLC的发生与发展。Objective To investigate the expression level of miR-196a in non-small cell lung cancer（NSCLC） tissues and cell lines,and to explore the effect of inhibition or up-regulation of miR-196a expression on NSCLC cells proliferation and its target gene.Methods Real-time quantitative PCR was performed to detect the relative expression of miR-196a in NSCLC cell lines and tissues.MiR-196a inhibitors or mimics were transfected into SPC-A1 or A549 cells to down-regulate or up-regulate miR-196a expression,and real-time quantitative PCR was used to test the transfection efficiency.MTT and colony formation assays were performed to detect the effect of miR-196a on NSCLC cells proliferation.Bioinformatics and Western blotting analysis were performed to investigate the target genes that could be regulated by miR-196a.Results This study showed that miR-196a was highly expressed both in NSCLC samples and cell lines compared with their corresponding normal tissues and cell lines.Transfection of miR-196a inhibitors or miR-196a mimics could significantly decrease or increase its＇ expression in SPC-A1 cells or A549 cells,respectively.Down-regulating of miR196a expression could inhibit SPC-A1 cells proliferation by inhibiting p27kip1 expression.Conclusion Up-regulated miR-196a could promote NSCLC cells proliferation,which may affect the development of NSCLC.

关 键 词：非小细胞肺癌 MIRNA miR-196a 细胞增殖 P27KIP1 

分 类 号：R734.2[医药卫生—肿瘤]