【摘要】 目的 探讨S100钙结合蛋白A4(S100A4)调控细胞周期蛋白D1(CCND1)转录本参与银屑病角质形成细胞异常增殖的作用。方法 2022年5月—2024年9月于新疆维吾尔自治区人民医院实验室进行实验。采用免疫组织化学法检测正常皮肤组织(23例)与寻常型银屑病患者(50例)皮损组织中CCND1蛋白表达；使用siRNA方式对S100A4进行沉默处理，高通量测序技术获得S100A4影响的转录组数据(RNA-seq),并对数据进行功能分析；在HaCaT细胞中使用S100A4的抗体，进行RNA紫外交联免疫共沉淀结合高通量测序(iRIP-seq),制备文库；RNA-seq检测siCtrl组和si-S100A4组中CCND1表达。结果 银屑病皮损组织CCND1蛋白阳性率为98%(49/50),高于正常皮肤组织的78.3%(18/23)(χ~2/P=7.947/0.005);对RNA-seq数据进行GO和KEGG分析，结果提示S100A4在调节细胞间信号、炎性反应、角质化、血管生成、细胞黏附、表皮发育中具有重要作用；将RNA-seq中获得的S100A4沉默后的差异表达基因与iRIP-seq 2次重复中鉴定到的S100A4结合峰基因进行overlap分析，发现重叠基因中包含CCND1基因；使用RNA-seq数据分析FPKM值发现，siCtrl组FPKM值(173.04±1.96)高于si-S100A4组的(54.72±1.27)(t/P=87.660/<0.001),提示沉默S100A4后CCND1基因表达下调。结论 S100A4结合CCND1,并影响其表达，从而影响细胞周期。两者共同作用参与银屑病的发生发展，影响角质形成细胞的增殖和凋亡。更多还原

【Abstract】 Objective To investigate the role of S100A4 in regulating CCND1 transcript to participate in abnormal proliferation of keratinocytes in psoriasis. Methods From May 2022 to September 2024, the experiment was conducted in the laboratory of People’s Hospital of Xinjiang Uygur Autonomous Region.Immunohistochemistry was used to detect the expression of CCND1 protein in normal skin tissues and lesional tissues of patients with plaque psoriasis. siRNA was applied to silence S100A4, and transcriptome data affected by S100A4 were obtained by high-throughput sequencing(RNA-seq) followed by functional analysis. In HaCaT cells, S100A4 antibody was used for RNA immunoprecipitation with high-throughput sequencing(iRIP-seq) to prepare libraries. RNA-seq was performed to detect CCND1 expression in siCtrl and si-S100A4 groups. Results The positive rate of CCND1 protein in lesional tissues was 98%(49/50), significantly higher than 78.3%(18/23) in normal skin tissues(χ~2/P=7.947/0.005). GO and KEGG analyses of RNA-seq data showed that S100A4 played important roles in regulating intercellular signaling, inflammatory response, keratinization, angiogenesis, cell adhesion, and epidermal development. Overlap analysis between differentially expressed genes after S100A4 silencing from RNA-seq and S100A4-binding peak genes identified in two iRIP-seq replicates revealed that CCND1 was included in the overlapping genes. FPKM value analysis from RNA-seq showed that the CCND1 expression was downregulated after S100A4 silencing, with a higher FPKM value in the siCtrl group than in the si-S100A4 group(P<0.001). Conclusion S100A4 binds to CCND1 and affects its expression, thereby influencing the cell cycle. Their combined action participates in the occurrence and development of psoriasis, affecting the proliferation and apoptosis of keratinocytes.更多还原