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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 the expression of CD44 in nasopharyngeal carcinoma and nasopharyngeal carcinoma cell lines by reverse transcriptase-polymerase chain reaction (RT-PCR) to find new human CD44 splice variant. METHODS: Specific primers were designed at both the start and stop codons of CD44 gene and at the splice site of exon 10 of variant splicing. RT-PCR was performed using nasopharyngeal carcinoma cells, cell lines 5-8F and HNE1 as templates, PCR amplification, product sequencing. Then, the cloned sequences were analyzed using bioinformatics. Results: The new cloned CD44 splice variant has 1634bp, contains a complete reading frame, the start codon in the sequence of 12, the stop codon in the sequence of 1301, the alternative splicing variant only spliced exon 10 , Predicted to encode 429 amino acids. GenBank accession number: EF581837. CONCLUSION: A novel CD44 splice variant predicted to encode 429 amino acids exists in human nasopharyngeal carcinoma tissues and cell lines studied, and its function needs further study.