De Felice Bruna, Garbi Corrado, Santoriello Margherita, Santillo Alessandra, Wilson Robert R
Department of Life Sciences, University of Naples II, Via Vivaldi 43, 81100 Caserta, Italy.
Mol Cell Biochem. 2009 Jul;327(1-2):191-201. doi: 10.1007/s11010-009-0057-x. Epub 2009 Feb 18.
Keloids are benign skin tumors and are the effect of a dysregulated wound-healing process in genetically predisposed patients. They are characterized by formation of excess scar tissue beyond the boundaries of the wound. Keloids are often confused with hypertrophic scars because of an apparent lack of morphologic differences. The molecular distinction between scars and keloid is still controversial and, until today, there is no appropriate treatment yet for keloid disease. In this study, we have found, for the first time, p53 mutations in both hypertrophic scar and keloids fibroblasts from cultured cells to various extents. Since p53 plays a central role in the DNA damage response by inducing cell cycle arrest and/or apoptotic cell death, we also set up time course experiments making cell cultures at different times to investigate the phenomenon of apoptosis and its involvement in the process of pathological scarring in both hypertrophic scars and keloids. The extent of apoptosis in this study was investigated by DNA fragmentation and MTT assays, propidium iodide staining, p53 expression, and subcellular distribution. Moreover, the correlation of apoptosis and ROS levels in keloid and hypertrophic scars fibroblasts was assessed. Understanding the molecular mechanisms that determine the regulation of apoptosis during wound healing might allow us to therapeutically modulate these pathways so that apoptotic cell death is reactivated in dysregulated and hypertrophic cells.
瘢痕疙瘩是良性皮肤肿瘤，是基因易感性患者伤口愈合过程失调的结果。其特征是在伤口边界之外形成过多的瘢痕组织。由于明显缺乏形态学差异，瘢痕疙瘩常与增生性瘢痕相混淆。瘢痕和瘢痕疙瘩之间的分子区别仍存在争议，直到今天，瘢痕疙瘩疾病仍没有合适的治疗方法。在本研究中，我们首次在不同程度上发现培养细胞来源的增生性瘢痕和成纤维细胞中的p53突变。由于p53通过诱导细胞周期停滞和/或凋亡性细胞死亡在DNA损伤反应中起核心作用，我们还进行了时间进程实验，在不同时间进行细胞培养，以研究凋亡现象及其在增生性瘢痕和瘢痕疙瘩病理瘢痕形成过程中的作用。本研究通过DNA片段化和MTT分析、碘化丙啶染色、p53表达和亚细胞分布来研究凋亡程度。此外，还评估了瘢痕疙瘩和成纤维细胞中凋亡与ROS水平的相关性。了解决定伤口愈合过程中凋亡调控的分子机制，可能使我们能够通过治疗调节这些途径，从而在失调和肥大细胞中重新激活凋亡性细胞死亡。
