紫外线对光化性角化病和正常皮肤p53、基质金属蛋白酶2、9表达的影响  

Effect of ultraviolet radiation on the expression of p53, matrix metalloproteinase-2 and -9 in actinickeratosis lesions and normal human skin

作　　者：徐丹[1,2] 刘彤云[1] 袁瑞红[1] 涂颖[1] 顾华[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》2013年第2期113-116,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30860257）

摘　　要：目的探讨不同剂量紫外线对光化性角化病（AK）与正常皮肤组织中增殖与凋亡、及p53、基质金属蛋白酶（MMP）2和MMP9表达的影响。方法AK与正常皮肤组织分别分为4个组，对照组（照射剂量为0J／cm2）、5J／cm2组、10J／cm2组、20J／cm2组。每块组织连续照射4d，继续培养24h后取材。TUNEL检测细胞凋亡、Ki～67检测细胞增殖情况，定量PCR及免疫组化法检测p53、MMP2及MMP9表达。结果紫外线照射后，与对照组相比，凋亡细胞百分比在正常皮肤组织10、20J／cm2组（46．8±2．1，56．7±2．4）增加，在AK20J／emz（43．5±1．5）组增加，正常皮肤组织10、20J／cm2组比AK同剂量组增加（P〈0．05）；Ki-67阳性细胞百分比在正常皮肤组织20J／cm2组（3．34±0．76）表达减少（P〈0．05），在AK中无明显变化（P〉0．05）；p53mRNA（5J／cm2：1．106±0．025；10J／cm2：1．259±0．045；20J／cm。：1．425±0．053）及蛋白（10J／cm2：0．1169±0．0032；20J／cm2：0．1454±0．0047）在正常皮肤组织中表达增加，在AK中，mRNA（10J／cm2：0．611±0．050；20J／cm2：0．578±0．070）及蛋白（20J／cm2：0．0404±0．0027）表达减少（P〈0．05）；正常皮肤组织中MMP2mRNA及蛋白（10J／cm2：1．086±0．013，0．0843±0．0024；20J／cm2：1．417±0．036，0．1236±0．0042）、MMP9mRNA及蛋白（20J／em0：1．395±0．026，0．3065±o．0162）表达增加，AK中MMP2mRNA及蛋白（20J／cm2：1．296±0．028，0．0744±0．0032）、MMP9mRNA及蛋白（10J／cm2：1．298±0．035，0．0992±0．0053；20J／cm2：1．286±0．032，0．1010±0．0063）表达增加（P〈0．05）。结论紫外线促进AK进展可能与其下调p53表达，上调MMP2和MMP9有关。Objective To estimate the effect of different doses of uhraviolet （UV） radiation on the proliferation of and apoptosis in kertatinocytes, as well as on the expression of p53, matrix metalloproteinase-2 （MMP2） and -9 （MMPg） in actinic keratosis（AK） lesions and normal human skin. Methods Tissue specimens were obtained from the lesions of 20 patients with AK and sun-exposed normal skin of 20 healthy human subjects, and subjected to an air-exposed culture. Each of the specimens was divided into 4 areas to remain untreated （control area） or be irradiated with UV of 5, 10 and 20 J/cm2 （irradiated areas） for 4 consecutive days. After another 24-hour culture, the tissue cultures were collected followed by the evaluation of apoptosis in and proliferation of keratinocytes by using terminal deoxynueleotidyl transferase-mediated dUTP nick end labeling （TUNEL） and Ki-67 staining, and determination of mRNA and protein expressions of p53, MMP2 and MMP9 by using real time PCR and immunohistochemistry respectively. Results A statistical increase was observed in the percentage of apoptotic cells in the normal skin irradiated with UV of 10 and 20 J/cm2 （46.8% ± 2.1% and 56.7% ± 2.4%, both P 〈 0.05） and in the AK lesions irradiated with UV of 20 J/cm2 （43.5% ± 1.5%, P 〈 0.05） compared with the corresponding unirradiated tissues. The normal skin showed a higher percentage of apoptotic cells than the lesional skin after irradiation with UV of 10 and 20 J/cm2 （both P 〈 0.05）. The percentage of Ki- 67-positive cells was significantly decreased in the normal skin after irradiation with UV of 20 J/cm2 （3.34% ± 0.76%, P 〈 0.05 ）, but experienced no statistical changes in the lesional skin after different doses of UV irradiation （all P 〉 0.05）. There was a statistical elevation in the expression of p53 mRNA （5 J/cm2：1.106 ± 0.025, 10 J/cm2：1.259 ± 0.045, 20 J/cm2：1.425 ± 0.053, all P 〈 0.05） and protein（ 10 J/cm2：0.1169 ± 0.0032, 20 J/cm2： 0.1454 ± 0.0047,

关 键 词：光化性角化病 紫外线 基因 P53 基质金属蛋白酶2 基质金属蛋白酶9 

分 类 号：R739.5[医药卫生—肿瘤]