CAJ | 학술논문

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mirVanaTM 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Feature Extraction( v10.7)软件对杂交图片进行分析,再用GeneSpring( GX10.0)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-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-1*等.其中上调基因hsa-miR-21和下调基因hsa-miR-451 miRNA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.
목적 탐토인증생성반흔여정상피부조직적미소RNA (miRNA)표체보차이.방법 수집2010년11월지2011년5월남창대학제일부속의원소상중심5례환자적증생성반흔급기린근정상피부조직,채용Trizol법추제총RNA,선용mirVanaTM miRNA분리시제합대기진행순화,연후사용미소RNA표기화잡교시제합진행형광표기급심편잡교,이용Feature Extraction( v10.7)연건대잡교도편진행분석,재용GeneSpring( GX10.0)연건진행수거귀일화급차이분석,동시응용실시정량반전록취합매련반응(RT-PCR)방법험증miRNAs심편결과적가고성.결과 사선출재인증생성반흔중표체상조적기인92개,표체하조적기인13개.기중현저상조적기인유hsa-miR-564,hsa-miR-936등；현저하조적기인유hsa-miR-451,hsa-miR-223,hsa-miR-363,hsa-miR-29b-1*등.기중상조기인hsa-miR-21화하조기인hsa-miR-451 miRNA적험증결과여검측결과구유교호적일치성.결론 인증생성반흔화정상피부조직중존재명현적miRNA차이성표체,가능여증생성반흔적발생、발전급연화밀절상관.
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 hyperplastic 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 (v10.7 ) software and the microarray results 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.