分泌型卷曲相关蛋白1在人毛乳头中的表达及其转录机制研究  

Expression and possible transcriptional mechanism of SFRP1 in the dermal papilla of human hair

作　　者：周蓝波 丁琦 曹倩[2] 冯丹丹 刘静静 徐宇轩 李志瑜 鲁严[1] 范卫新[1] ZHOU Lan-bo;DING Qi;CAO Qian;FENG Dan-dan;LIU Jing-jing;XU Yu-xuan;LI Zhi-yu;LU Yan;FAN Wei-xin(Department of Dermatology and Venereology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,Chinn)

机构地区：[1]南京医科大学第一附属医院皮肤性病科,江苏南京210029 [2]南京医科大学第一附属医院儿科,江苏南京210029

出　　处：《临床皮肤科杂志》2021年第1期12-16,共5页Journal of Clinical Dermatology

基　　金：国家自然科学基金(81972954)资助项目。

摘　　要：目的:探讨人分泌型卷曲相关蛋白1(secreted frizzled related protein 1, SFRP1)基因在人毛乳头中的表达及其转录机制。方法:应用免疫组化观察SFRP1在雄激素性秃发(androgenetic alopecia, AGA)患者秃发区的毛囊与非AGA患者的正常毛囊毛乳头部位的表达情况。采用按基因组序列设计的引物,通过PCR克隆人SFRP1启动子片段,将之插入荧光素酶报告基因载体pGL3-basic中,构建不同截短片段的重组质粒并瞬时转染毛乳头细胞,测定质粒在毛乳头细胞中的启动子活性,找到核心启动子区域,运用生物信息学方法预测其潜在的转录因子结合位点。结果:免疫组化结果显示AGA患者毛乳头中SFRP1表达量显著高于非AGA患者。经菌液PCR、双酶切、测序,成功构建了人SFRP1启动子及其截短片段重组质粒,并确定其启动子活性区域位于-100~+50bp区域内,且可能含有E2F3、HIF1A::ARNT、FOXC1、SP1等转录因子结合位点。结论:SFRP1在AGA患者毛乳头中表达呈阳性,人SFRP1启动子荧光素酶报告质粒构建成功,其转录因子结合位点初步确定,为进一步的研究奠定基础。Objective: To investigate the expression of human secreted frizzled related protein 1(SFRP1) in the dermal papilla of human hair and its transcriptional mechanism. Methods: Expression of SFRP1 in the dermal papilla of hair from androgenetic alopecia(AGA) patients and non-AGA patients was observed by immunohistochemistry. Original human promoter fragments were cloned by PCR, using primers generated from genomic sequences of SFRP1. Amplified fragments were then cloned into the pGL3-basic vector. Additional three recombinant plasmids of different lengths obtained through deletion were cloned into pGL3-basic vectors, which were then transfected into dermal papilla cells, respectively. The expression of each vector was determined by measuring luciferase activity in dermal papilla cells. The potential transcriptional binding sites were predicted by bioinformatics methods. Results: The immunohistochemical findings showed that the expression of SFRP1 in the dermal papilla of AGA patients was much higher than that in non-AGA patients. The human SFRP1 promoter and its truncated fragment recombinant plasmids were successfully constructed, and the core promoter region was determined to be located in the -100~+50 bp region, which may contain the binding sites of E2F3, HIF1A::ARNT, FOXC1, SP1 and other transcription factors. Conclusion: SFRP1 is expressed in the dermal papilla of AGA patients. Recombinant plasmid of human SFRP1 promoter has been successfully constructed, and the possible transcription factor binding sites have been identified. Our data provide a basis for further study.

关 键 词：分泌型卷曲相关蛋白1(SFRP1) 毛乳头 转录机制 

分 类 号：R758.71[医药卫生—皮肤病学与性病学]