MicroRNAs在瘢痕疙瘩中的差异表达  

Differential expression of microRNAs in human keloids

作　　者：郭晓瑞 梁杰[2] 黄如林[3] 卢玲[2] 金玉丹 罗少军[2] 吴志远[2] 

机构地区：[1]中山市第二人民医院激光美容整形中心,广东省中山市528447 [2]广东医学院附属医院整形外科研究所,广东省湛江市524001 [3]上海交通大学医学院附属第九人民医院整复外科,上海市200011

出　　处：《中国组织工程研究》2012年第50期9370-9375,共6页Chinese Journal of Tissue Engineering Research

基　　金：广东省科技计划项目基金项目(2010B031600286)资助~~

摘　　要：背景:瘢痕疙瘩的形成受多种基因、细胞因子的调控,然而目前对这些基因、细胞因子的调控表达机制的研究尚不明确。目的:通过检测人正常皮肤组织和人瘢痕疙瘩组织中microRNAs的表达情况,初步筛选人瘢痕疙瘩microRNAs表达谱。方法:根据瘢痕疙瘩临床诊断标准及实验要求制定瘢痕疙瘩、正常皮肤病例纳入标准和排除标准,收集瘢痕疙瘩标本(6例)和正常皮肤标本(5例);TRIZOL法提取标本的总RNAs并进行质检,再采用Ambion's miRNA Isolation Kit从总RNAs中分离microRNAs;荧光标记后与微阵列芯片杂交,扫描检测后数据经SAM和Cluster分析处理,筛选瘢痕疙瘩组织中microRNA差异表达谱。采用实时荧光定量RT-PCR技术验证在瘢痕疙瘩组织中均存在显著差异表达的microRNAs。结果与结论:通过microRNA微阵列芯片筛选得到人瘢痕疙microRNAs差异表达谱(筛选出显著差异表达microRNAs12个),与正常皮肤组织比较,microRNAs在瘢痕疙瘩组织中表达下调。提示瘢痕疙瘩microRNAs差异表达可能参与瘢痕疙瘩的发生发展过程,部分microRNAs有可能成为用于瘢痕疙瘩诊治的专有分子标志物。BACKGROUND：Keloid formation is affected by regulations of many genes and cytokines.However,it is not clear about the mechanism underlying the regulation of these genes and cytokines expressions.OBJECTIVE：To investigate the differential expression of microRNAs between human normal skins and keloid tissues,and to initially screen microRNAs expression profiling in keloid tissues.METHODS：According to clinical diagnostic criteria and experiment demands,the inclusion criteria and exclusion criteria of keloids and normal skins were formulated：keloids（six cases） and normal human skins（five cases）.Total RNAs were extracted from keloids and normal skin tissues by Trizol,and microRNAs were further isolated by Ambion＇s miRNA Isolation Kit.In keloids,the differential expression profile of microRNAs was harvested by significance analysis of microarrays（SAM） and Cluster,based on microarray screening.Real time quantitative reverse transcription-PCR was applied to verify reliability of microRNA array results.RESULTS AND CONCLUSION：Differential expression profile of MicroRNAs（n=12） was obtained in human keloids,which may be related to the pathogenesis and development of keloids.Moreover,part of them may be used for unique molecular markers for diagnosis and treatment of keloids.

关 键 词：瘢痕疙瘩 MICRORNAS 瘢痕 microRNA微阵列芯片 实时荧光定量RT-PCR 差异表达 分层聚类 基因 调控 荧光标记 

分 类 号：R318[医药卫生—生物医学工程]