微小RNA-135b在胃癌发生发展中的临床病理意义及其作用  

Clinicopathological significance and function of miR-135b in the occurrence and development of gastric cancer

作　　者：王霖沛[1] 马筱秋[1] 蔡建春[1] 

机构地区：[1]厦门大学附属第一医院厦门市肿瘤中心肿瘤外科,361003

出　　处：《中华医学杂志》2012年第46期3269-3273,共5页National Medical Journal of China

基　　金：国家自然科学基金（81172283）

摘　　要：目的探讨微小RNA-135b（miR-135b）在胃癌发生发展中的作用及其临床病理意义。方法收集厦门大学附属第一医院肿瘤外科自2007年9月至2011年3月72例胃癌组织及其相应胃正常上皮组织标本，选取其中6例采用微小RNA（miRNA）微阵列技术检测胃癌miRNA表达谱并筛选差异表达的miRNA。以miR-135b作为研究对象，用实时定量（RT）-PCR检测其在上述临床标本、正常胃上皮细胞系GES-1、胃癌细胞系BGC-823和SGC-7901中的表达水平。采用细胞转染、噻唑蓝（MTT）法、流式细胞技术研究miR-135b对胃癌细胞增殖、细胞周期及凋亡的作用。应用RT—PCR和蛋白质印迹探讨miR．135b与结肠腺瘤性息肉病基因（APC）之间的关系。结果在胃癌组织中，表达上调和下调超过2倍的miRNA分别为7个和9个（均P〈0．05）。与各自相应的胃正常上皮组织相比，miR-135b在绝大多数胃癌组织中的表达水平更高（66例，91．67％）。在所有临床标本中，二者间的平均表达量差异有统计学意义（3．42±2．62和1．00±0．07，P〈0．01），并且与多种胃癌病理参数如Laur6n分型、分化程度、浸润深度和pTNM分期密切相关（均P〈0．05），在分化越差的细胞系中其表达水平越高（P〈0．05）。miR-135b能促进胃癌细胞增殖，抑制其凋亡，并靶向调控APC在胃癌细胞中的表达。结论胃癌组织具有自己独特的miRNA表达谱。miR-135b的高表达对胃癌的发生发展起重要作用，它参与对APC的调控可能是胃癌发病机制之一。Objective To explore the function and clinicopathological significance of miR-135b in the occurrence and development of gastric cancer. Methods Seventy-two pairs of fresh gastric cancer tissues and corresponding normal gastric epithelium were collected at our department from September 2007 to March 2011. Six of them were randomly selected and miRNA microarray was applied to study the miRNA expression profiles of gastric cancer. Quantitative real-time PCR was used to validate the reliability of mieroarray and detect miR-135b expression in the above clinical samples, as well as cell lines GES-1, BGC-823 and SGC- 7901. The methods of cell transfection, thiazolyl blue tetrazolium bromide （MqT） and flow cytometry were used to study the impact of miR-135 on cell proliferation, cell cycle and apoptosis of gastric cancer cells. Real-time quantitative PCR and Western blot were used to explore the relationship between miR-135b and adenomatous polyposis coli （APC）. Results Compared with normal gastric tissues, 7 miRNA were significantly up-regulated and 9 miRNA significantly down-regulated for over 2 folds in gastric cancer tissues （P〈0. 05）. The results of microarray were verified by quantitative real-time PCR. MiR-135b expression was up-regulated in most clinical samples compared with their corresponding epithelium （n = 66, 91.67% ）. The average expression level of miR-135b in gastric cancer tissues was significantly higher than normal epithelium （3.42 ＋2. 62 vs 1.00 -＋0. 07, P 〈0. 05）. MiR-135b was related to Laurrn classification, tumor differentiation, invasion and pathologic tumor-node-metastasis （pTNM） stage of gastric cancer （all P 〈 0. 05 ）. The worse differentiation degree of gastric cell lines, the higher miR-135b expression level （P 〈 0. 05 ）. MiR-135b promoted gastric cancer cell proliferation, inhibited its apoptosis and directly regulated the expression of APC in gastric cancer cell. Conclusions Special miRNA expression profiles of gastric cancer are found. MiR-135b

关 键 词：微RNAS 微阵列分析 胃肿瘤 基因 APC miR-135b 

分 类 号：R735.2[医药卫生—肿瘤]