摘要
目的:应用逆转录多聚酶链式反应(RT-PCR)分析CD44在鼻咽癌组织和鼻咽癌细胞株中表达,来寻找新的人CD44剪接变异体。方法:在CD44基因起始和终止密码子两端及变异剪接外显子v10中和剪接位点处设计特异性引物,以鼻咽癌组织、细胞株5-8F和HNE1 cDNA为模板进行RT-PCR扩增,产物测序。然后,应用生物信息学对克隆序列进行分析。结果:新克隆的CD44剪接变异体有1634bp,包含一个完整的阅读框,起始密码子在序列的12位,终止密码子在序列的1301位,可变剪接区只有变异型剪接外显子10,预测编码429氨基酸。GenBank登陆号:EF581837。结论:一个新的、预测编码429个氨基酸的CD44剪接变异体存在于研究的人鼻咽癌组织和细胞株中,它的功能有待于进一步研究。
AIM： To detect unknown CD44 variants （CD44v） in nasopharyngeal cancer by using reverse transeription - polymerase chain reaction （ RT - PCR） to analyze the expression of cell adhesion protein CD44 gene in nasopharyngeal cancer tissue and cell lines. METHODS： Specific primers at up start code, down terminal code of CD44 and primers at the middle, splicing points of variable splicing exon v10 of CD44 were designed, eDNA of nasopharyngeal cancer tissues, 5 - 8F and HNE1 cell lines were analyzed by RT - PCR. Products of RT - PCR were sequenced and further analyzed by bioinformaties. RESULTS ： The new CD44v sequence possessed 1634 bp, with a completed open reading frame. The start code was at 12 bp site and terminal code at 1301bp site. It was predicted to code 429 amino acids, and only variable splicing exon 10 existed in the flexible region. It was given an accessible number EF581837 by GenBank. CONCLUSION： A new CD4 variant predicted to code 429 amino acids exists in the studied nasopharyngeal cancer tissues and cell lines.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第9期1734-1737,共4页
Chinese Journal of Pathophysiology
基金
广西壮族自治区卫生厅项目(No.Z2007236)
广西壮族自治区科技厅科技攻关项目(No.0592007-1J)
广西高校病理学人才小高地建设创新团队基金资助项目(No.桂教人[2005]34)
关键词
CD44
剪接变异体
鼻咽肿瘤
CD44
Spliced isoform
Nasopharyngeal neoplasms
