ALA-PDT对皮肤鳞状细胞癌A431细胞甲基化及Gadd45a表达的影响  

Experimental Study of Topical 5-aminolevulinic Acid-photodynamic Therapy on Methylation and Expression of Gadd45a in Squamous Cell Carcinoma of Skin Cell Line A431

作　　者：李小静[1] 李志锋[1] 李显平[1] 王久江[1] 靳玉荣[1] 李海滨[1] 燕霞[1] 刘保国[1] 刘志军[1] L! Xiaojing;LI Zhifeng;LI Xianping;WANG Jiujiang;JIN Yurong;LI Haibin;YAN Xia;LIU Baoguo;LIU Zhijun(Department of Dermatology ,Affilaited Hospital of Hebei University of Engineering, Handan 056002, Chin)

机构地区：[1]河北工程大学附属医院皮肤科,河北邯郸056002

出　　处：《中国皮肤性病学杂志》2018年第6期615-619,共5页The Chinese Journal of Dermatovenereology

基　　金：河北省科技计划项目(15277721D)

摘　　要：目的探究5-氨基酮戊酸光动力疗法(ALA-PDT)对皮肤鳞状细胞癌(简称皮肤鳞癌)A431细胞甲基化及Gadd45a表达的影响。方法培养皮肤鳞癌A431细胞,将细胞分为对照组以及不同剂量的ALA-PDT处理组(ALA浓度分别为:0.1、0.5、2mmol/L),MTT检测各组细胞增殖活力。分别采用q PCR和Western blot方法检测ALA-PDT处理后A431细胞Gadd45a mRNA及Gadd45a蛋白表达。流式细胞术检测ALA-PDT处理后各组细胞凋亡率,并进一步观察经ALA-PDT处理后对A431细胞DNA总体甲基化水平的影响。结果不同ALA浓度(0.1、0.5、2mmol/L)ALA-PDT处理后,随ALA浓度上升A431细胞增殖抑制率及细胞凋亡率均呈逐渐升高的趋势(P均<0.05);经不同ALA浓度(0.1、0.5、2mmol/L)ALAPDT处理后A431细胞Gadd45a蛋白表达水平随ALA浓度上升而上升,而ALAPDT组DNA总体甲基化显著下降,与对照组相比差异有统计学意义(P<0.05)。结论 ALA-PDT治疗通过促进Gadd45a蛋白表达抑制皮肤鳞癌A431细胞增殖并促进凋亡,过表达的Gadd45a蛋白可抑制A431细胞DNA甲基化。Objective To investigate the effects of topical 5-aminolevulinic acid-photodynamic therapy（ALA-PDT）on proliferation, apoptosis, the expression of growth arrest and DNA-damage-inducible protein 45a（Gadd45a）, and methylation in squamous cell carcinoma of skin cell line A431. Methods Cultured A431 cells were transfected, and classified into the control group, 0.1mmol/L ALA, 0.5mmol/L ALA, and 2mmol/L ALA groups. Cell viability was detected by MTT assay. The mRNA and protein levels of Gadd45a were evaluated by quantitative real-time polymerase chain reaction（qRT-PCR）and Western blotting respectively. The rate of cell apoptosis was detected by flow cytometry in each group, and the DNA methylation level was further determined. Results The protein level of Gadd45a was elevated with the increase of ALA concentration（P〈0.05）. The inhibition rate of A431 cell proliferation as well as apoptosis was positively associated with ALA concentration（P〈0.05）. Compared with the control group, the DNA methylation level notably reduced in the ALA-PDT group（P〈0.05）. The DNA methylation level showed a negative correlation with the ALA concentration（P〈0.05）. Conclusion ALA-PDT inhibits A431 cell proliferation and promotes cell apoptosis, and in the meantime ALA-PDT suppresses the DNA methylation by accelerating the protein level of Gadd45a.

关 键 词：ALA—PDT Gadd45a蛋白 皮肤鳞状细胞癌 甲基化 增殖分化 

分 类 号：R739.5[医药卫生—肿瘤]