Department of Dermatology, Wakayama Medical University, Wakayama, Japan.
Department of Plastic Surgery, Wakayama Medical University, Wakayama, Japan.
J Dermatol. 2023 Sep;50(9):1170-1179. doi: 10.1111/1346-8138.16861. Epub 2023 Jun 27.
Hypertrophic scars and keloids are fibroproliferative disorders caused by abnormal wound healing. Their exact cause has not been found, but abnormalities during the wound healing process including inflammatory, immune, genetic, and other factors are thought to predispose an individual to excessive scarring. In the present study, we performed transcriptome analysis of established keloid cell lines (KEL FIB), focusing on gene expression analysis and fusion gene detection for the first time. For gene expression analysis, fragments per kilobase per million map read values were calculated, which were validated by real-time PCR and immunohistochemistry. Fusion genes were predicted by transcriptome sequence, and validated by Sanger sequence and G-banding. As a result, GPM6A was shown in the expression analysis to be upregulated in KEL FIB compared with normal fibroblasts. The GPM6A upregulation in KEL FIB was confirmed by real-time PCR, and GPM6A messenger ribonucleic acid expression was consistently significantly elevated in the tissues of hypertrophic scar and keloid compared to normal skin. Immunohistochemistry also revealed that the number of fibroblast-like spindle-shaped cells positive for GPM6A was significantly increased in keloidal tissues. GPM6A inhibition by small interfering ribonucleic acid significantly reduced the number of KEL FIB. On the other hand, although we hypothesized that fusion genes are involved in the pathogenesis of keloids, the transcriptome analysis could not prove the presence of fusion genes in KEL FIB. Taken together, GPM6A upregulation may have an inducible effect on cell proliferation in keloidal fibroblasts. GPM6A can be a novel therapeutic target in hypertrophic scars and keloids. The inflammatory nature may be more prominent in the pathogenesis of keloids, rather than being skin tumors, as proposed by Ogawa et al. Future studies using several cell lines will be required.
增生性瘢痕和瘢痕疙瘩是由异常伤口愈合引起的纤维增生性疾病。其确切病因尚未发现，但认为伤口愈合过程中的异常包括炎症、免疫、遗传和其他因素，使个体容易产生过度瘢痕形成。在本研究中，我们首次对已建立的瘢痕疙瘩细胞系（KEL FIB）进行了转录组分析，重点进行基因表达分析和融合基因检测。对于基因表达分析，计算了每百万映射读数片段的每千碱基数（fragments per kilobase per million map read values），并通过实时 PCR 和免疫组织化学进行了验证。通过转录组序列预测融合基因，并通过 Sanger 序列和 G 带验证。结果表明，与正常成纤维细胞相比，KEL FIB 中的 GPM6A 在表达分析中上调。实时 PCR 证实了 KEL FIB 中的 GPM6A 上调，与正常皮肤相比，增生性瘢痕和瘢痕疙瘩组织中的 GPM6A 信使核糖核酸表达始终显著升高。免疫组织化学还显示，GPM6A 阳性的成纤维细胞样梭形细胞数量在瘢痕疙瘩组织中显著增加。小干扰核糖核酸抑制 GPM6A 显著减少 KEL FIB 的数量。另一方面，尽管我们假设融合基因参与瘢痕疙瘩的发病机制，但转录组分析不能证明 KEL FIB 中存在融合基因。总之，GPM6A 的上调可能对瘢痕疙瘩成纤维细胞的细胞增殖具有诱导作用。GPM6A 可以成为增生性瘢痕和瘢痕疙瘩的新治疗靶点。Ogawa 等人提出，瘢痕疙瘩的发病机制可能更具炎症性质，而不是皮肤肿瘤。需要使用几种细胞系进行进一步的研究。
