瘢痕疙瘩旁真皮成纤维细胞转录组特征初探  

Transcriptomic characteristics of keloid-adjacent dermal fibroblasts:a preliminary study

作　　者：张道宁 林萍萍 田杰 张国红 李航[1] Zhang Daoning;Lin Pingping;Tian Jie;Zhang Guohong;Li Hang(Department of Dermatology and Venereology,Peking University First Hospital,National Clinical Research Center for Skin and Immune Diseases,Beijing Key Laboratory of Molecular Diagnosis on Dermatoses,NMPA Key Laboratory for Quality Control and Evaluation of Cosmetics,Beijing 100034,China;Department of Pathology,Shantou University Medical College,Shantou 515041,Guangdong,China)

机构地区：[1]北京大学第一医院皮肤性病科,皮肤病分子诊断北京市重点实验室,国家皮肤与免疫疾病临床医学研究中心,国家药品监督管理局化妆品质量控制与评价重点实验室,北京100034 [2]汕头大学医学院病理学系,汕头515041

出　　处：《中华皮肤科杂志》2025年第2期145-153,共9页Chinese Journal of Dermatology

摘　　要：目的从转录组水平描述瘢痕疙瘩的潜在皮损范围,为解析瘢痕疙瘩切除后复发提供分子证据。方法收集2022年7-12月在北京大学第一医院皮肤性病科临床确诊并经手术切除治疗的瘢痕疙瘩患者3例,分别取其瘢痕疙瘩及瘢痕疙瘩旁真皮组织,同时收集4例良性皮肤肿瘤旁真皮组织作为对照。通过原代细胞培养纯化获得真皮成纤维细胞,传至第2代后进行转录组测序并进行差异基因筛选、基因本体论(GO)功能以及京都基因和基因组数据库(KEGG)通路富集分析。在分析差异表达基因时,定义差异表达倍数(FC)>2且P<0.05的基因为表达上调基因,FC<0.5且P<0.05的基因为表达下调基因。通过分析已发表文献中瘢痕疙瘩与瘢痕疙瘩旁组织的单细胞转录组测序数据(GSA数据库中HRA000425)及正常真皮单细胞转录组测序数据(GEO数据库中GSE130973),侧面验证本研究中转录组结果的准确性。在本研究及文献中的组织样本中验证瘢痕疙瘩旁关键基因的表达。结果与瘢痕疙瘩成纤维细胞相比,瘢痕疙瘩旁及对照真皮成纤维细胞共同的表达上调基因63个,富集在脂质运输(P=0.038)、离子运输(P=0.040)等生物学过程;与对照真皮成纤维细胞相比,瘢痕疙瘩旁及瘢痕疙瘩成纤维细胞共同的表达上调基因56个,富集在转化生长因子β信号通路(P<0.001)等。瘢痕疙瘩、瘢痕疙瘩旁真皮成纤维细胞分别与对照真皮成纤维细胞相比,仅在瘢痕疙瘩旁表达上调的差异基因有79个;经基因表达基值过滤与一致性分析,最终获得与瘢痕疙瘩旁转录组特征密切相关的候选基因13个(平均表达量>1000且组内表达量方差<30000的基因),包括抑制瘢痕疙瘩形成相关基因SMAD6、SMAD7及促进瘢痕疙瘩形成相关基因MSX1、SNAI1、EDN1,同样参与细胞生长、骨化、软骨形成等生物学过程(均P<0.01)。通过ChEA3网站进行转录因子的富集分析发现,上述13个基因富集Objective To explore the potential lesional range of keloids by analyzing the transcriptomic characteristics,and to provide a molecular basis for understanding the recurrence of keloids following surgical excision.Methods From July to December in 2022,3 patients clinically diagnosed with keloids and treated with surgical excision at the Department of Dermatology and Venereology,Peking University First Hospital were included in the study.Samples of keloids and keloid-adjacent dermis were collected from these 3 patients,and normal dermal tissues adjacent to benign skin tumors were collected from 4 patients and served as controls.Dermal fibroblasts were obtained by primary cell culture and purification,which were then subsequently passaged to the second generation for transcriptome sequencing.Differential gene expression analysis,gene ontology(GO)-based functional analysis,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed.While analyzing differential expression genes,those with a fold change(FC)>2 and a P value<0.05 were defined as upregulated genes,whereas those with a FC<0.5 and a P value<0.05 were considered downregulated genes.The accuracy of the results was further validated by comparing them with published single-cell sequencing data on keloid and keloid-adjacent tissues(HRA000425 in GSA database)and single-cell sequencing data on the normal dermis(GSE130973 in GEO database).Key genes in keloid-adjacent dermal fibroblasts were validated in tissue samples from this study and the literature.Results Compared with keloid-derived fibroblasts,keloid-adjacent and control dermal fibroblasts shared 63 upregulated genes enriched in biological processes including lipid transport(P=0.038)and ion transport(P=0.040);compared with control dermal fibroblasts,keloid-adjacent and keloid-derived fibroblasts shared 56 upregulated genes enriched in the transforming growth factorβsignaling pathway(P<0.001),etc.When comparing keloid-derived fibroblasts and keloid-adjacent fibroblasts with c

关 键 词：瘢痕疙瘩 转录组 成纤维细胞 差异表达基因 瘢痕疙瘩旁真皮组织 

分 类 号：R622[医药卫生—整形外科]