摘要
目的探讨人增生性瘢痕与正常皮肤组织的微小RNA（miRNA）表达谱差异。方法收集2010年11月至2011年5月南昌大学第一附属医院烧伤中心5例患者的增生性瘢痕及其邻近正常皮肤组织，采用Trizol法抽提总RNA，先用mirVana TM miRNA分离试剂盒对其进行纯化，然后使用微小RNA标记和杂交试剂盒进行荧光标记及芯片杂交，利用FeatureExtraction（v10．7）软件对杂交图片进行分析，再用GeneSpring（GXl0．0）软件进行数据归一化及差异分析，同时应用实时定量反转录聚合酶链反应（RT．PCR）方法验证miRNAs芯片结果的可靠性。结果筛选出在人增生性瘢痕中表达上调的基因92个，表达下调的基因13个。其中显著上调的基因有hsa-miR一564，hsa—miR-936等；显著下调的基因有hsa．miR-451，hsa．miR-223，hsa-miR-363，hsa—miR-29b一1}等。其中上调基因hsa-miR-21和下调基因hsa．miR-451miRNA的验证结果与检测结果具有较好的一致性。结论人增生性瘢痕和正常皮肤组织中存在明显的miRNA差异性表达，可能与增生性瘢痕的发生、发展及演化密切相关。
Objective To explore the miRNA differential expression profiles of hyperplastic scar and normal skin so as to further elucidate the pathogenesis of hyperplastic scar and search for new theraputic targets. Methods The total RNA was extracted from 5 human byperplastic scar and normal skin tissues by Trizol. The specimens were collected from the First Affiliated Hospital of Nanchang University from November 2010 to May 2011, and purified by mirVanaTM miRNA Isolation Kit and then labeled and hybridized by miRNA Complete Labeling and Hyb Kit. The images of hybridization were analyzed by the Feature Extraction （ vl0. 7 ） software and the microarray resuhs confirmed by quantitative real-time reverse transcription-polymerase chain reaction （RT-PCR）. Results In hyperplastic scar, 92 miRNA genes were up-regulated and 13 down-regulated. The most significantly up-regulated miRNAs were hsa-miR-564 and hsa-miR-936, etc. while hsa-miR-451, hsa-miR-223, hsa-miR-363 and hsa-miR-29b-1 ＊ became significantly down-regulated. The findings of RT-PCR on hsa-miR-21 and hsa-miR-451 ofregulation were in a high concordance with the microarray results. Conclusion Distinct differences of miRNA expression between human hyperplastic scar and normal skin, it may be closely correlated with the formation, development and evolution of hyperplastic scar.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2012年第10期692-694,共3页
National Medical Journal of China
基金
国家自然科学基金（81060323）
关键词
瘢痕
微RNAS
基因表达谱
Cicatrix
MicroRNAs
Gene expression profiling
