光化性角化病转化生长因子β1／Smads调控p53表达的研究  

Regulation of p53 expression though transforming growth factorβ1 （TGFβ1）/Smads pathway in actinic keratosis

作　　者：徐丹[1,2] 袁瑞红[1] 涂颖[1] 汤諹[1] 顾华[1] 张丽[1] 何黎[1] 

机构地区：[1]昆明医科大学第一附属医院皮肤科,650032 [2]昆明医科大学基础医学院

出　　处：《中华皮肤科杂志》2012年第9期638-640,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30860257）

摘　　要：目的通过光化性角化病（AK）皮肤组织块干扰模型探讨转化生长因子（TGF）β1／Smads对p53的调控作用。方法培养人AK组织块，分为5个组，第1组对照组，第2组TGFβ1培养24h组，第3组SB431542培养24h组，第4组TGFβ1培养48h组，第5组SB431542培养48h组，取材后实时PCR和Western印迹分别检测p53mRNA和蛋白表达及Smad2磷酸化水平。结果在TGFβ1培养24h后，与对照相比，p53mRNA表达增加（13．4968±0．9903，P〈0．05），Smad2磷酸化增多（0．700±0．023，P〈0．05）；培养48h后，p53mRNA为13．38824-1．6772，蛋白为1．009±0．001，均增加（P〈0．05），Smad2磷酸化增多（0．646±0．120，P〈0．05）；TGFβ1培养24h和48h相比，p53蛋白由0．634±0．040增加至1．009±0．001（P〈0．05）；在SB431542培养24h后，与对照相比Smad2磷酸化变化不明显，当SB431542培养48h后，与对照相比Smad2磷酸化水平明显减少（0．116±0．003，P〈0．05），其余没有明显变化。结论人AK皮肤组织块培养可作为一种信号通路的干扰模型，TGFβ1在AK中可通过Smads通路调控p53表达。Objective To evaluate the performance of actinic keratosis （AK） tissue as a culture model for the study of interference in transduction pathway, and to explore the mechanism underlying the p53 regulation though TGFβ1/Smads pathway by using the tissue culture model. Methods Twenty-five skin samples from the lesions of patients with AK were cultured, and divided into 5 groups to be treated with TGFβ1 of 10μg/L for 24 and 48 hours, the transforming growth factor （TGF） β1 receptor kinase inhibitor SB431542 of 10 μmol/L for 24 and 48 hours, respectively, or remain untreated. Real time PCR and Western blot were performed to quantify the mRNA expression of p53 and protein expression of p53 and phosphorylated Smad2 in these tissue specimens respectively. Results A significant elevation was observed in the expressions of p53 mRNA （13.4968 ± 0.9903 vs. 1, P 〈 0.05） and phosphorylated Smad2 （0.700 ± 0.023 vs. 1, P 〈 0.05） in AK tissues after treatment with TGFβ1 for 24 hours, and in the expressions of p53 mRNA （13.3882 ± 1.6772 vs. 1, P 〈 0.05） and protein （1.009 ± 0.001 vs. 0.512 ± 0.005, P 〈 0.05） after treatment with TGFβ1 for 48 hours, compared with the untreated AK tissues. No significant differences were observed in the expression of p53 protein between the AK tissues treated with TGFβ1 for 24 hours and 48 hours （P 〉 0.05）. SB431542 induced a statistical reduction in the level of phosphorylated Smad2 at 48 hours （0.116 ± 0.003 vs. 0.306 ± 0.023, P 〈 0.05）, but no significant changes were observed in the expression of p53 mRNA or protein after SB431542 treatment for 24 or 48 hours. Conclusions AK tissue cultures can serve as a model for the study of interference in signal transduction pathway. TGFβ1 might regulate the expression of p53 protien through Smads pathway in AK.

关 键 词：光化性角化病 转化生长因子Β1 Smad2蛋白质 基因 P53 

分 类 号：R739.5[医药卫生—肿瘤]