目的探讨E3泛素连接酶WWP1、p53在人正常皮肤及瘢痕疙瘩组织中的表达差异,分析其与瘢痕疙瘩发生是否相关。方法选取临床切取的瘢痕疙瘩及正常皮肤组织标本各10例,采用免疫组织化学法和RT—PCR法分别对组织中WWP1、p53蛋白及mRNA表达情况进行检测,并通过Image—Pro Plus图像分析系统测定组织蛋白染色后累计光密度值、以B—actin为参照计算mRNA相对定量,进而比较分析两组标本中WWP1、p53表达是否有差异性。结果免疫组化染色结果提示,瘢痕疙瘩标本中p53蛋白表达明显低于正常皮肤,而WWP1蛋白沉积明显高于正常皮肤,同时p53及WWP1 mRNA RT-PCR检测证实同样结果,差异均有统计学意义(P〈0.05)。结论E3泛素连接酶WWP1及抑癌基因p53与瘢痕疙瘩的形成有相关性,可作为瘢痕疙瘩形成机制及治疗靶点研究的方向。
Objective The purpose of the study was to detect the expression of E3 ubiquitin ligase WWP1 and p53 in human normal skin and keloid tissues and its correlation with the occurrence of keloid. Methods 10 cases of keloid tissues and 10 cases of normal skin were recruited. The expressions of WWP1 and p53 in the keloid group and the controls were measured using immunohistochemical methods. And the mRNA expression of WWP1 and p53 in the two groups were detected by RT-PCR methods. We also measured integrated optical density (1OD) of the proteins by Image-Pro Plus analysis system, and calculated the relative quantity of mRNA refering to 13-actin, then made a comparative analysis of the two groups for the expressions of WWP1, p53. Results The immunohistochemical results showed that the expression of p53 protein in keloid was significantly lower compared with the controls, and WWP1 protein deposition was significantly higher than that of normal skin. The mRNA expression of p53 and WWP1 was detected by RT-PCR and confirmed the same results. All the difference was statistically significant (P 〈0.05). Conclusion E3 ubiquitin ligase WWP1 and anti oneogene p53 is related to the formation of keloid, which provides a new direction of the mechanism and new targets for the treatment of keloid.