苯并蒽诱导金黄地鼠颊黏膜癌变的重要致病基因筛选  

Screening of virulence gene in golden hamster cheek poch mucosa carcinomatous change induced by 9, 10-dimethylen-1,2-benzanthracene

作　　者：张国栋[1] 杨凯[1] 梅杰[1] 

机构地区：[1]重庆医科大学附属第一医院口腔颌面外科,400016

出　　处：《中华口腔医学杂志》2010年第5期303-306,共4页Chinese Journal of Stomatology

基　　金：重庆市医学科技计划（2006）

摘　　要：目的 利用基囚芯片和生物信息学分析技术探讨口腔黏膜癌变发生发展的不同阶段基因表达谱的改变，并筛选癌变的重要致病基因。方法用9，10-二甲基1，2苯并蒽（9，10-dimethylen-1，2-benzanthracene，DMBA）诱导建立金黄地鼠颊鳞状细胞痛模型，分别提取并纯化正常颊黏膜、癌前病变和鳞状细胞痛组织的总RNA并合成用Cy3荧光标记的cRNA，分别与含有41000条基因-表达序列标签（expressedsequence tags，EST）的Agilent大鼠全基因表达谱芯片杂交，以Ratio≥2和≤0．5为阈值确定癌变3个不同阶段的差异表达基因并分析相关生物信息，然后用GeneSpnng10．0软件行Venn图分析，筛选小在口腔颊黏膜癌变发生发展的3个不同阶段均持续表达异常的基因，用反转录聚合酶链反应（RT—PCR）对部分差异表达基凶行验证分析。结果金黄地鼠颊黏膜自正常黏膜到鳞状细胞癌的过程中，共有5255条差异表达基因，其中表达上调2896条，下调2359条。癌变的3个不同发展阶段均持续表达异常的基因共有22条，其中表达上调3条，下调19条。对上调基因EaG2和下调基因Ecg-2行RT—PCR验证结果与芯片结果相符。结论口腔黏膜癌变涉及众多基因表达的改变；癌变的不同阶段均持续表达异常的基因可能为癌变的重要致病基因，对以上基因的探讨将对研究口腔黏膜癌变发生发展的分子机制和靶向治疗具有重要作用。Objective To examine and analyze the global gene expression at the different stages of golden hamster cheek poch mucosa carcinomatous change induced by 9, 10-dimethylen-1,2 benzanthracene （DMBA）. Methods The model of golden hamster cheek poch squamous cell carcinoma was induced by DMBA. The RNA of normal mucosa, precancerous lesions and squamous cell carcinoma of fresh tissue of golden hamsters was extracted and purified and the cRNA labeled by fluorescent Cy3 synthesized, which respectively hybridized with the agilent rat cDNA microarray containing 41000 genes-expressed sequence tags,scanning with Agilent G2565AA fluorescence scanner. The Ratio ≥2 and Ratio ≤0. 5 were used to determine the threshold value of the differences gene expression and analyze the relevant biological information of the three different stages of cancer. The Venn diagram by Gene Spring 10. 0 analysis software was then used to select the genes of continuing expression in the three different stages of cancer. Meanwhile, RT-PCR was used to confirm the correctness of the results of the gene chip. Results A total of 5255 differentially expressed genes were detected during the process from normal mucosa to the squamous cell carcinoma,of which 2896 was up-regulated and 2359 down-regulated. There were 22 genes that showed continues abnormal expression through the three different stages of carcinomatous change, including 3 upregulated, 19 down-regulated. The RT-PCR results of Ear-2 and Ecg-2 were consistent with the gene chip. Conclusions The development of oral mucosal squamous cell carcinoma involved a number of abnormal genes. The genes showing continues abnormal expression at different stages of carcinomatous change may be the important pathogenetic ones.

关 键 词：口腔黏膜 癌 鳞状细胞 基因 

分 类 号：R739.8[医药卫生—肿瘤]