摘要
目的:利用全基因芯片技术筛选口腔颊黏膜癌前病变发生发展的相关致病基因。方法:用9、10-二甲基;1,2-苯并蒽(Dimethyl benzanthracene,DMBA)诱导建立金黄地鼠颊鳞癌模型,分别提取正常颊黏膜和癌前病变组织的总RNA并合成用Cy3荧光标记的cRNA,分别与含有41000个基因/ESTs序列的Agilent大鼠全基因芯片杂交,以Log2Ratio≥2.5和Log2Ratio≤-2.5为阈值来确定差异表达基因,然后用Gene Ontology对差异表达基因行功能分类分析,最后用RT-PCR对部分差异表达基因进行验证。结果:金黄地鼠颊黏膜自正常黏膜到癌前病变发展过程中,共得到1331条差异表达基因,其中上调基因747条,下调基因584条。Gene Ontology分析发现这些差异基因主要涉及到大分子代谢、信号传导等8大功能分类。对上调基因Atp6v0d2和下调基因Sfrp2行RT-PCR验证结果与芯片结果相符。结论:口腔颊黏膜癌前病变的发生涉及众多基因表达的改变,通过对这些基因的进一步研究,将对探索口腔黏膜癌前病变的发病机制、有效的干预防止癌前病变的发生或逆转癌前病变具有重要意义。
Objective：The aim of this study was to screen and analysize the important virulent genes in golden hamster cheek pochmucosa with precancerous lesions induced by DMBA.Methods：The models of golden hamster cheek poch mucosa precancerous lesions were induced by DMBA（styrene-dimethyl-anthracene）.We extracted the total RNA from the normal mucosa and precancerous lesions;then we respectively used the Cy3 to mark the cDNA of normal mucosa and precancerous lesions;and used the Agilent Rat microarray containing 41,000 genes/ESTs to screen the differentially expressed genes;used the screening criteria of Log2Ratio ≥2.5 and Log2Ratio≤-2.5,at last using the Gene Ontology to analysis the biology function of the important virulent genes.Meanwhile,we confirmed the correctness of the results of the Gene Chip by using the RT-PCR to verificate some important pathogenic genes.Results：We detected a total of 1331 differentially expressed genes（up-regulated 747;down-regulated 584）.GO analysis showed that these differentially expressed genes mainly related to the macromolecular metabolism,signal transduction and so on.The RT-PCR results of Atp6v0d2 and Sfrp2 were consistent with the gene chip analysis.Conclusion：Towards the screening and analysis of the important differentlily expressed genes in the gold hamster cheek pouch mucosa with precancerous lesions,we found that the special stage-precancerous lesions involved in changes in multiple genes.The further study of these genes is useful to explore the pathogenesis of precancerous lesions in the oral mucosa and effectively prevent the occurrence of precancerous lesions.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2010年第5期706-710,共5页
Journal of Chongqing Medical University
基金
重庆市卫生局科研项目(编号:03-2-073)
关键词
颊囊黏膜
癌前病变
全基因表达谱
基因芯片
Cheek poch mucosa
Precancerous lesions
Gene expression profiling
Gene chip
