E Yang, Qipa Zou, Hengshu Zhang
Wounds. 2014 May;26(5):139-46.
This study aimed to investigate the expression and significance of DNA methyltransferase 1 (DNMT1) in pathologic scar fibroblasts, as well as the influence of methylase inhibitor, 5-aza-2-deoxycytidine, on pathological scars.
Samples of 31 keloids, 20 hypertrophic scars, and 25 normal skins were taken to test the expression rate of DNMT1 by immunohistochemistry. Primary fibroblasts were cultured with the monoplast method. Samples were categorized into the keloid group (K group), 5-aza-2-deoxycytidine keloid intervention group (K+ group), normal skin group (N group), hypertrophic scar group (H group), and 5-aza-2-deoxycytidine hypertrophic scar intervention group (H+ group). The expressions of DNMT1, transforming growth factor-β (TGF-β), and Smad7 mRNA in each group were detected with realtime polymerase chain reaction. The effect of 5-aza-2-deoxycytidine on the cell cycle and apoptosis of pathologic scar fibroblasts were analyzed with flow cytometry.
The expression rate of DNMT1 was 100% in keloid fibroblasts, 90% in hyperplastic scar fibroblasts, and 8% in normal skin fibroblasts. After the intervention with 5-aza-2-deoxycytidine in the K+ group, the expression of DNMT1 and TGF-β1 mRNA was lower, Smad7 mRNA was elevated in pathological scar fibroblasts, the flow cytometry showed the proportion of cells in G0/G1 phase were increased, and the proportion of apoptosis cells were also increased, with similar changes in the cells in the H and H+ groups.
DNMT1 may play a vital role on the generation of pathological scars. Methylaze inhibitors 5-aza-2-deoxycytidine may influence the related cytokines of pathological scars, inhibit proliferation, and promote apoptosis of pathological scar fibroblasts. The generation of pathological scars may be related with methylation of certain genes. 5-aza-2-deoxycytidine be a new choice for the treatment of pathological scars.
本研究旨在探讨DNA甲基转移酶1（DNMT1）在病理性瘢痕成纤维细胞中的表达及意义，以及甲基化酶抑制剂5-氮杂-2'-脱氧胞苷对病理性瘢痕的影响。
取31例瘢痕疙瘩、20例增生性瘢痕及25例正常皮肤标本，采用免疫组织化学法检测DNMT1的表达率。采用单细胞法培养原代成纤维细胞。样本分为瘢痕疙瘩组（K组）、5-氮杂-2'-脱氧胞苷瘢痕疙瘩干预组（K+组）、正常皮肤组（N组）、增生性瘢痕组（H组）和5-氮杂-2'-脱氧胞苷增生性瘢痕干预组（H+组）。采用实时聚合酶链反应检测各组中DNMT1、转化生长因子-β（TGF-β）和Smad7 mRNA的表达。采用流式细胞术分析5-氮杂-2'-脱氧胞苷对病理性瘢痕成纤维细胞细胞周期及凋亡的影响。
瘢痕疙瘩成纤维细胞中DNMT1的表达率为100%，增生性瘢痕成纤维细胞中为90%，正常皮肤成纤维细胞中为8%。K+组经5-氮杂-2'-脱氧胞苷干预后，病理性瘢痕成纤维细胞中DNMT1和TGF-β1 mRNA的表达降低，Smad7 mRNA升高，流式细胞术显示G0/G1期细胞比例增加，凋亡细胞比例也增加，H组和H+组细胞也有类似变化。
DNMT1可能在病理性瘢痕的形成中起重要作用。甲基化酶抑制剂5-氮杂-2'-脱氧胞苷可能影响病理性瘢痕的相关细胞因子，抑制病理性瘢痕成纤维细胞的增殖并促进其凋亡。病理性瘢痕的形成可能与某些基因的甲基化有关。5-氮杂-2'-脱氧胞苷有望成为病理性瘢痕治疗的新选择。
