喉癌差异表达cDNA序列的分离与初步鉴定  

Isolation and Identification of cDNA Sequences Differentially Expressed in Laryngeal Carcinoma

作　　者：李友军[1] 田芳[1] 陈主初[1] 关勇军[1] 何春梅[1] 杨新明[2] 谢鼎华[2] 

机构地区：[1]湖南医科大学肿瘤研究所,长沙410078 [2]湖南医科大学附属第二医院耳鼻喉科,长沙410007

出　　处：《生物化学与生物物理学报》2000年第2期153-157,共5页

基　　金：国家自然科学基金!(No :3 990 0 0 5 2 );卫生部科学研究基金!(No :98 118)资助项目&&

摘　　要：分离和克隆人喉癌中新的相关基因将有助于揭示喉癌的易感性与癌变机制。运用mRNA差异显示法对 2例成人喉癌组织及配对癌旁正常组织的基因表达进行研究 ,分离到 3 5个差异显示片段 ;用反向Northern点杂交筛选到 6个差异片段 ,经克隆、测序和匹配分析 ,得 12条不同cDNA序列 ,其中 4条为新基因序列 ,另外 8条与已知基因高度同源。将 12条cDNA序列固定在膜上 ,用来自喉癌和配对正常组织的总cDNA探针与其杂交和差异RT PCR分析 ,鉴定了它们在喉癌组织和癌旁正常组织中差异表达。上述结果提示这些差异cDNA序列可能与喉癌发病密切相关。The isolation of the genes related to laryngeal carcinoma(LC) is necessary for revealing mechanisms of carcinogenesis and genetic predisposition to LC. The mRNA differential display method was used to compare and analyze mRNAs prepared from two adult laryngeal carcinoma tissues and paired tumor adjacent normal tissues. A total of twenty two differential display experiments was performed and thirty five cDNA fragments differentially expressed in normal or malignant laryngeal epithelial tissues were identified. Differential expression of six of these thirty five cDNA fragmens was confirmed by reverse northern dot blot. Subsequent cloning of six differentially expressed cDNA fragments and sequencing and BLASTN analysis resulted in the identification of twelve distinct cDNA sequences. Four of these were shown to be novel gene sequences that had not been reported. Eight of the remaining cDNA sequences showed sequence homology to those previously reported. The differential expression of these twelve cDNA sequences in the carcinoma or normal tissue of the larynx were confirmed by fixing the twelve cDNA sequences on the membrane, followed by the hybridization with the total cDNA probes from laryngeal carcinoma or normal tumor adjacent tissues and by differential RT PCR. These results suggested that these cDNA sequences might be involved in carcinogenesis of laryngeal carcinoma.

关 键 词：喉癌 MRNA差异显示 差异表达 发病原因 

分 类 号：R739.65[医药卫生—肿瘤]