miRNA-24下调S100A8蛋白表达能够抑制人喉鳞癌Hep2细胞侵袭  

miRNA-24 inhibits human laryngeal squamous carcinoma Hep2 cells invasion by regulating S100A8 protein expression

作　　者：郭艳[1] 富伟能[2] 陈洪[2] 尚超[2] 

机构地区：[1]中国医科大学口腔医学院中心实验室,辽宁沈阳110007 [2]中国医科大学基础医学院医学遗传学教研室,辽宁沈阳110001

出　　处：《现代肿瘤医学》2011年第12期2389-2393,共5页Journal of Modern Oncology

基　　金：中国医科大学口腔医学院青年科研启动基金资助项目(编号:K101593-11-28)

摘　　要：目的:探讨microRNA-24(miRNA-24)对喉鳞状细胞癌(laryngeal squamous carcinoma,LSCC)Hep2细胞侵袭影响的分子机制。方法:应用miRanda和RNA22靶基因预测软件预测miR-24的靶基因及其结合位点,根据预测结果通过转染miR-24前体上调其在Hep2细胞中的表达,应用荧光定量PCR和Western blot分别检测miR-24过表达对S100钙结合蛋白A8(S100 calcium binding protein A8,S100A8)mRNA和蛋白表达变化的影响。S100A8抗体阻断方法,检测转染miR-24前体后Hep2细胞侵袭能力的变化。结果:S100A83'非翻译区(3'-untranslation region,3'UTR)含有一个miR-24结合位点,而且该位点与大鼠的同源性高达95%;miR-24基因转染的Hep2细胞中miR-24的表达显著升高,S100 A8蛋白的表达显著降低,(P均<0.05),而S100A8 mRNA表达变化未见显著性差异,(P>0.05)。与未阻断组相比,miR-24前体转染能明显降低阻断S100A8组的Hep2细胞侵袭能力,(P<0.05)。结论:LSCC Hep2细胞中,miR-24可结合到S100A8基因的3'UTR,在转录后水平上负性调控S100A8基因的表达,从而抑制Hep2细胞侵袭。Objective：To explore the molecular mechanism of microRNA-24（miR-24） on laryngeal squamous carcinoma（LSCC） Hep2 cells invasion ability.Methods： Software miRanda and RNA22 were used to predict the miR-24 targeted gene and its binding sites,and then quantitative real-time PCR and Western blot were used to detect the expression of S100A8 mRNA and protein respectively after transfection of Hep2 cells with pre-miR-24 according to the prediction result.The invasive ability of Hep2 cells were assayed by Transwell after anti-S100A8 blocked.Results： S100A8 3＇UTR contained a miR-24 binding site,which had a homology up to 95% with rat.In transfected cells,the increased accumulation of miR-24 resulted in a significantly decreased S100A8 protein level（P〈0.05）,but had no effect on the level of S100A8 mRNA（P〈0.05）.The invasion ability of Hep2 cells were statistically inhibited while compared anti-S100A8 blocked group with non-blocking group,P〈0.05.Conclusions： miR-24 could negatively regulate the expression of S100A8 by binding to the 3＇UTR,which lead LSCC Hep2 cells invasion ability inhibited.

关 键 词：喉鳞癌 microRNA-24 S100A8 侵袭 

分 类 号：R730.23[医药卫生—肿瘤] R739.65[医药卫生—临床医学]