肺癌差异表达cDNA序列的分离与鉴定  

Isolation and Identification of cDNA Sequence Differentially Expressed in Human Lung Carcinoma

作　　者：谢海龙[1] 李友军[1] 陈主初[1] 关勇军[1] 何春梅[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所细胞生物研究室,湖南长沙410078

出　　处：《癌症》2002年第3期249-253,共5页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.39900052)

摘　　要：背景与目的:肺癌癌变的分子机理至今还不很清楚,其关键是未找到与肺癌密切相关的基因。本研究拟分离和鉴定肺癌差异表达cDNA序列,为进一步克隆肺癌相关基因打下基础。方法:采用mRNA差异显示、克隆、反向点杂交、测序和Reversetranscription-PCR(RT-PCR)方法分离在肺癌组织和肺癌细胞系中表达下调或不表达,以及在肺癌组织和肺癌细胞系中过表达的差异cDNA序列。结果:mRNA差异显示获得16条差异的片段。将差异片段进行克隆、反向点杂交、测序、同源性比较,共获得10条不同的cDNA序列。其中2条为新基因,8条与已知基因高度同源。进一步RT-PCR证实LXDD1、LXDD2两条cDNA序列在肺癌及正常肺支气管粘膜组织中存在差异表达。结论:运用mRNA差异显示,成功分离了10条差异表达cDNA序列,并证实其在肺癌及正常肺支气管粘膜组织中存在表达差异。这些差异表达cDNA序列可能与肺癌的发生发展密切相关。Background &Objective:The molecular mechanism of the pathogenesis of lung carcinoma is unclear;because key genes related to lung carcinoma have not been identified.Thi s study was designed to isolate and id entify differentially expressed cDNA sequences in human lung carcinoma for clo ning lung carcinoma-related genes.Method:Using mRNA differential display method combined with cloning and reserve Northern dot blot and sequencing and RT-P CR,differentially expressed cDNA sequences were isolated between two lung cancer cell lines ,two adult lung carcinoma tissue and paired normal tissue of trachea e pithelia.Result:Sixteen differentially expressed c DNA fragments were isolated,subsequent cloning of sixteen diffe rentially expressed cDNA fragments were confirmed by reverse Northern d ot blot and sequencing and BLAST analysis.Two o f these were shown to be novel gene seq uences that had not been reported.Ei ght of the remaining cDNA sequences homolo gize to known genes;additionally tw o differentially expressed cDNA seq uences were confirmed by RT-PCR.Conclusion:Using mRNA differentially display,The authors had successfully isolated ten differentially expressed cDNA sequences from human lung carcinoma,these cDNA sequences might be involved i n the pathogenesis of lung carcinoma.

关 键 词：肺肿瘤 MRNA差异显示 差异表达 CDNA 序列分析 

分 类 号：R734.2[医药卫生—肿瘤]