生物钟基因Perl和细胞周期基因p53、CyclinD1、B1、CDK在金黄地鼠口腔颊黏膜癌变不同阶段昼夜节律表达变化及与癌变发生的相关性分析  

A comparative study on the variation in expression levels of Perl and cell cycle gene p53,Cyclin D1,B1,CDK of Circadian gene in different stages of oral mucosa carcinogenesis and its correlation with carcinogenesis

作　　者：全知怎[1] 陈盈哲 汪大林[1] 

机构地区：[1]上海第二军医大学第一附属医院口腔科,上海200433

出　　处：《临床和实验医学杂志》2017年第18期1772-1776,共5页Journal of Clinical and Experimental Medicine

摘　　要：目的探讨生物钟基因Perl和细胞周期基因p53、Cyclin D1、B1、CDK在金黄地鼠口腔颊黏膜癌变不同阶段昼夜节律表达变化及与癌变发生的相关性。方法在黑暗和光照交替环境下饲养90只金黄地鼠,在地鼠的颊黏膜上涂抹二甲基苯并蒽(DMBA)以建立颊黏膜鳞状细胞癌模型,分别在涂抹DMBA前和涂抹DMBA6周、14周后的24小时开灯后时间(HALO)的第4个HALO开始,每隔4HALO处死5只金黄地鼠,分别获取正常颊黏膜和癌前病变以及鳞状细胞癌3个阶段各昼夜6个时间点的组织。按照常规将获取的组织进行切片,在HE染色下对组织的癌变情况进行观察;对各时间点组织中p53、Cyclin D1和B1、CDK1 mRNA、Perl等表达水平采用定时定量荧光PCR(RT-PCR)进行检测。结果 Perl mRNA在鳞状细胞癌组和癌前病变组以及正常颊黏膜组中各时间点的表达均有明显差异(P<0.05);鳞状细胞癌组和癌前病变组的Perl mRNA表达中值及振幅均显著低于正常颊黏膜组(P<0.05),且鳞状细胞癌组又显著低于癌前病变组(P<0.05)。细胞周期基因p53、细胞周期蛋白D1和周期蛋白依赖性激酶的mRNA在三组各时间点之间的表达差异均有统计学意义(P<0.05),而细胞周期蛋白B1的mRNA则仅在鳞状细胞癌组和正常颊黏膜组各时间点中的表达差异具有统计学意义(P<0.05)。鳞状细胞癌组和癌前病变组的p53 mRNA表达中值均显著低于正常颊黏膜组(P<0.05),且鳞状细胞癌组又显著低于癌前病变组(P<0.05)。鳞状细胞癌组和癌前病变组的CDK1 mRNA表达中值均显著高于正常颊黏膜组(P<0.05),鳞状细胞癌组的Cyclin B1 mRNA表达中值显著高于正常颊黏膜组(P<0.05)。结论金黄地鼠实验结果提示,生物钟基因Perl和细胞周期基因p53、Cyclin D1、B1、CDK的昼夜节律均随着口腔颊黏膜癌变的发生发展而变化,即细胞周期和昼夜节律之间的相互作用与癌变的发生具有极为密切的关系。Objective To explore the biological clock gene Perl and cyclin dependent kinase（ CDK）,cell cycle protein（ Cyclin） D1,B1,cell cycle gene expression of p53 in oral buccal mucosa cancer in different stages of circadian rhythm changes and correlation with carcinogenesis. Methods A total of 90 golden hamsters were feeding in dark and light enviroment alternately,and anthracene（ dimethylbenzanthracene,DMBA） was pasted to the buccal mucosa of hamsters to establish models of buccal squamous cell carcinoma,the time before the paste of DMBA and 6 and 14 weeks after the paste,24 hours on time after light onset（ HALO）,5 golden hamsters were killed every 4 HALO,specimens were obtained from normal mucosa,precancerous lesions and squamous cell carcinoma of these 3 stages at 6 time points in the whole day. According to the conventional slicing,carcinogenesis was observed by HE staining; and the expression levels of CDK1 mRNA,Cyclin D1 and B1,p53,Perl were determined by quantitative fluorescence PCR（ real time PCR,RT-PCR）. Results The expression of Perl mRNA in squamous cell carcinoma group,precancerous lesion group and normal buccal mucosa group at each time point had obvious difference（ P〈0. 05）. The expression level and amplititude of Perl mRNA were significantly lower than those of normal buccal mucosa group（ P〈0. 05）,and they in squamous cell carcinoma group were significantly lower than those of precancerous lesion group（ P〈0. 05）. Cell cycle gene p53 protein and cell cycle D1 protein kinase were dependent and the difference in cycle of mRNA expression among various time points in three groups was statistically signficant（ P〈0. 05）,while the difference in expression of cell cycle protein B1 mRNA in squamous cell carcinoma group and normal buccal mucosa group in all time points was statistically significant（ P〈0. 05）. The level of p53 mRNA in squamous cell carcinoma group and precancerous lesion group was significantly lower than that of normal buccal mucosa group（ P〈0. 0

关 键 词：金黄地鼠 口腔颊黏膜 癌变 生物钟基因 细胞周期基因 昼夜节律 

分 类 号：R739.85[医药卫生—肿瘤]