老年皮肤血管瘤特征及microRNA调节的机制研究

Study on Features of Cutaneous Hemangioma in Elderly Patients and the MicroRNA Regulation Mechanism

目的从microRNA水平上寻找影响血管瘤血管发生的因素,并结合其下游蛋白的表达情况分析其与血管瘤发生的相关性。方法收集老年血管瘤,血管肉瘤等各种血管异常的标本,HE染色组织学观察;实时PCR分析microRNA表达;免疫组织化学检测α-SMA和IV型胶原;免疫荧光检测MEK-1、CYCLIN-E1同CD34共同表达的情况。原位杂交法检测microRNA-96。培养血管内皮细胞株并转染MEK-1和CYCLIN-E1 siRNAs验证蛋白表达的趋势。结果 microRNA PCR分析表明老年血管瘤组织microRNA-96水平比其他血管异常组织低。microRNA-96目的基因MEK-1和CYCLIN-E1蛋白表达较正常皮肤和其他血管异常组织增加。转染MEK-1和CYCLIN-E1的siRNA后,内皮细胞数量下降。结论老年血管瘤microRNA-96表达的减少和MEK-1、CYCLIN-E1水平的增加可以导致细胞异常增殖。

Objective To investigate the risk facts that affect angiogenesis in hemangioma on microRNA level,and analyze the relationship between the expression of downstream proteins and the hemangioma development.Methods Venous abnormities tissue was collected and histologic analyzed by HE staining.The microRNA expressions were checked by real time PCR.α-SMA and IV collagen were detected by immunohistochemistry.The co-expression of MEK-1,CYCLIN-E1 and CD34 were tested with immunofluorescence method.The microRNA-96 was localized by in situ hybridization.Meanwhile,siRNAs of MEK-1 and CYCLIN-E1 were introduced into the vascular endothelial cells and the expression trend of the protein was verified in vitro.Results microRNA PCR analysis showed the level of microRNA-96 was lower in senile hemangioma.The target genes MEK-1 and CYCLIN-E1 of microRNA-96 were increased compared with other tissues.After transfection of MEK-1 and CYCLIN-E1 siRNAs,the cells numbers were decreased.Conclusion The decreased expression of microRNA-96 and increased expression of MEK-1,CYCLIN-E1 in senile hemangioma could lead to abnormal cell proliferation.