Department of Environmental, Biological and Pharmaceutical Sciences and Technologies, University of Campania Luigi Vanvitelli, I‑81100 Caserta, Italy.
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II‑Medical School, I‑80131 Naples, Italy.
Mol Med Rep. 2018 May;17(5):7081-7088. doi: 10.3892/mmr.2018.8749. Epub 2018 Mar 15.
Keloids are benign skin tumors that develop in individuals who have a positive family history of keloid disorders. Keloids are characterized by a deregulated wound‑healing process, atypical fibroblasts with extreme deposition of extracellular matrix components, particularly collagen, increased cell proliferation and associated failure of apoptosis. Recently ingenol‑mebutate has been used as a novel agent with anti‑proliferative activity on human keloids as an alternative treatment option in patients, once conventional therapies have failed. We hypothesized that microRNAs (miR/miRNA) may be involved in the balance between lesion formation and repair. A comprehensive understanding of the molecular mechanism underlying the Ingenol‑mebutate response in keloid fibroblast following Ingenol‑mebutate exposure has been established previously. Therefore, the present study analyzed changes in miRNAs and apoptotic gene regulation in Ingenol‑mebutate treated keloid fibroblast, by reverse transcription‑quantitative polymerase chain reaction and a DNA fragmentation assay. The range of upregulated miRNAs and downregulated genes encoding cell death appeared to be associated with the degree of the morphological alterations in Ingenol‑mebutate treated keloids. In particular, the upregulation of miR‑34a was detected in keloid fibroblasts during and following Ingenol‑mebutate exposure. Keloid fibroblasts that overexpressed miR‑34a showed differential expression of genes involved in the apoptotic signaling pathway such as p53. In conclusion, the Ingenol‑mebutate treatment used here was effective in reducing keloid fibroblast growth in cell culture experiments and the expression of particular miRNAs modulated the pro‑apoptotic gene expression following Ingenol-mebutate treatment.
瘢痕疙瘩是一种良性皮肤肿瘤,发生于有瘢痕疙瘩家族病史的个体。瘢痕疙瘩的特征是伤口愈合过程失调、成纤维细胞异常,细胞外基质成分(尤其是胶原)过度沉积、细胞增殖增加以及细胞凋亡失败。最近, ingenol ‑ mebutate 已被用作一种新型药物,对人类瘢痕疙瘩具有抗增殖活性,作为传统治疗失败后患者的替代治疗选择。我们假设 microRNAs(miR/miRNA)可能参与病变形成和修复之间的平衡。之前已经建立了 Ingenol ‑ mebutate 暴露后 Ingenol ‑ mebutate 处理的瘢痕疙瘩成纤维细胞中 Ingenol ‑ mebutate 反应的分子机制的全面理解。因此,本研究通过逆转录‑定量聚合酶链反应和 DNA 片段化分析,分析了 Ingenol ‑ mebutate 处理的瘢痕疙瘩成纤维细胞中 miRNA 和凋亡基因调控的变化。上调的 miRNA 和下调的编码细胞死亡的基因的范围似乎与 Ingenol ‑ mebutate 处理的瘢痕疙瘩的形态改变程度相关。特别是,在 Ingenol ‑ mebutate 暴露期间和之后,检测到瘢痕疙瘩成纤维细胞中 miR ‑ 34a 的上调。过表达 miR ‑ 34a 的瘢痕疙瘩成纤维细胞显示参与凋亡信号通路的基因表达的差异,如 p53。总之,这里使用的 Ingenol ‑ mebutate 治疗在细胞培养实验中有效减少了瘢痕疙瘩成纤维细胞的生长,并且特定 miRNA 的表达调节了 Ingenol-mebutate 处理后的促凋亡基因表达。