皮肤鳞状细胞癌组织中p16及14-3-3σ基因启动子甲基化检测及其相关性分析  

Analyses of p16 and 14-3-3σ promoter methylation, and their association with skin squamous cell carcinoma

作　　者：齐淑贞 李静 刘浥 刘维达 

机构地区：[1]中国医学科学院北京协和医学院皮肤病医院性病科,江苏南京210042

出　　处：《临床皮肤科杂志》2017年第12期835-839,共5页Journal of Clinical Dermatology

基　　金：若干皮肤病性病省内诊疗体系建设与转化医学研究基金(BL2012007)资助项目

摘　　要：目的:通过检测皮肤鳞状细胞癌组织及正常皮肤组织中p16及14-3-3σ基因启动子甲基化,探讨p16及14-3-3σ基因启动子甲基化在皮肤鳞状细胞癌发生发展中的作用。方法:实时荧光定量聚合酶链反应(q-PCR)法检测53例皮肤鳞状细胞癌组织及48例正常皮肤组织中p16和14-3-3σm RNA表达,免疫印迹(western blot)法检测p16和14-3-3σ蛋白表达,应用甲基化特异性PCR对所提DNA进行p16及14-3-3σ基因启动子甲基化检测,并分析这两个基因的相关性。结果:皮肤鳞状细胞癌标本中p16及14-3-3σm RNA表达均低于正常皮肤组织,皮肤鳞状细胞癌标本中p16和14-3-3σ蛋白亦低于正常皮肤组织,皮肤鳞状细胞癌标本中p16及14-3-3σ基因启动子甲基化的阳性率分别为54.72%(29/53)和67.92%(36/53),高于对照组的8.33%(4/48)和18.75(9/48),差异有统计学意义(P<0.001);皮肤鳞状细胞癌组织中p16和14-3-3σ的甲基化存在正相关关系(r=0.431,χ~2=9.825,P=0.002,P<0.05)。结论:皮肤鳞状细胞癌标本中p16及14-3-3σ低表达及甲基化与鳞状细胞癌发生发展可能有关,皮肤鳞状细胞癌基因启动子甲基化可能是多位点参与的。Objective： To compare p16 and 14-3-3σ gene promoter methylation in skin squamous cell carcinoma （SCC） with that in normal skin, and to investigate pathogenic roles of these methylations in squamous cell carcinoma. Methods： SCC specimens from 53 patients and normal skin tissues from 48 individuals were used to assess the expression levels of p16 and 14-3-3σ proteins and mRNA, using Western blot and real-time fluorescence-based quantitative PCR, respectively. Methylation-specific PCR was used to assess promoter methylation of p16 and 14-3-3σ genes. Correlations between p16 or 14-3-3σ methylation and clinicopathological parameters were analyzed with the Spearman rank test. Results： SCC specimens showed significantly lower levels of both p16/14-3-3σ mRNA and protein compared to normal skin. In contrast, promoter methylation rates of p16and 14-3-3σ were significantly higher in SCC than in normal skin（p16：54.72% vs. 8.33%; 14-3- 3σ： 67.92% vs. 18.75%, both P〈0.001）. Methylation rates of p16 and 14-3-3σ genes in SCCs displayed positive correlation （r=0.431, X^2=9.825, P=0.002）. Conclusions： The low expression levels, and methylations of p16 and 14-3-3σ genes may play pathogenic roles in SCC, and gene promoter methylations at multiple sites may be involved in SCC.

关 键 词：皮肤鳞状细胞癌 P16基因启动子 14—3—3σ基因启动子 甲基化 

分 类 号：R739.5[医药卫生—肿瘤]