Different Expression And Significance Of WWP-1 And P53 In Keloid And Normal Skin

Background and Objective: Keloid is a manifestation of pathological skin repair.It has similar biological characteristics with tumor,and a certain genetic predisposition,by the interaction of multiple genes and many factors. In recent years,ubiquitination process was more and more deeply involved in a variety of tumors. E3 ubiquitin ligase is an enzyme that specifically recognizes the substrate in the ubiquitination process, which may be related to the mechanism involved in tumor about proliferation aging and signal transduction pathway. In 2010,a Japanese scholar found NEDD4-1genes of E3 ubiquitin ligase may be related to keloid by GWAS. E3 ubiquitin ligaseWWP1(The WW domain containing E3 ubiquitin protein ligase 1), as one of NEDD4 family protein,also has HECT and WW domains.The abnormal expression of WWP1 has been demonstrated in a variety of tumors, including breast and prostate cancer is the detailed report types. P53 gene is a recognized tumor suppressor gene, located on chromosome 17q13.1,involved in many tumor pathogenesis like apoptosis and cell cycle regulation, DNA repair, tumor development process. Keloid,characterized by the invasive growth of fibroblasts cell, abnormal proliferation and apoptosis and extracellular matrix deposition,may be related to the loss of some gene function about apoptosis control. The experiments detected the WWP1 and P53 gene and expression by immunohistochemistry and RT-PCR of keloid and normal skin tissue, and then explore their significance in keloid formation.Materials and Methods: We selected 10 keloid cases, aged 10 to 52 years old(33.1± 9.7), 6 males and 4 females.10 cases of normal skin specimens were also selected,aged 10 to 52 years old(31.8 ± 7.4).These specimens were divided into two parts,and half of them were done in PCR analysis part, detecting m RNA of WWP1 and P53;the other part as immunohistochemistry, in detecting protein of WWP1 and P53. We taked out keloid tissue and normal skin tissue stored in the-80 ° C liquid nitrogen into the homogenizer with TRIzol RNA extract, extracted total RNA and detected concentration and purity of the RNA using UV spectrophotometer.Then reverse transcription and amplification. Another part of the organization by sections,dewaxing, the antibody is added and incubated, color, were mounted after dehydration was observed under a microscope and photographed. Using Image-Pro Plus image analysis system to measure the transfer of E3 ubiquitin enzyme and accumulation of P53 protein optical density(Integrated Optical Density, IOD) value.Results: By staining two groups of P53 protein we found rare positive cells and significantly reduced the IOD value of stained keloid tissues P53 protein, and the difference was statistically significant(P <0.05). The IOD value of stained keloid E3 ubiquitin ligase WWP1 increased significantly and the difference was statistically significant(P <0.05). Compared with normal skin, the m RNA level of keloid E3 ubiquitin ligase WWP1 significantly increased, while the expression of P53 m RNA is significantly reduced compared to the control group, the differences were statistically significant(P <0.05).Conclusions:By this study,we found the decrease expression of P53 but increase expression of WWP1 in keloid,illustrating loss function of P53 in keloid. In our experiments of WWP1 level testing, we found significantly increased expression,and presumably WWP1 Ubiquitinated P53 protein, thereby forming a keloid fibroblasts proliferation associated with loss function of P53, whereas WWP1 regulate P53 to control of the cell cycle.