应用抑制消减杂交技术克隆鼻咽癌转移调控基因

Cloning of metastasis-associated genes in nasopharyngeal carcinoma using suppression subtractive hybridization

目的:分离并鉴定高低转移表型不同的人鼻咽癌细胞株差异表达cDNA序列,了解鼻咽癌转移抑制基因或具有抑制活性的核苷酸序列。方法:以具有高低转移潜能不同的人鼻咽癌细胞株为研究对象,应用抑制消减杂交技术(suppression sub tractivehybrid ization,SSH)和T/A克隆法构建差异表达cDNA消减文库,对差异表达的cDNA进行测序和生物信息学分析。结果:成功构建人鼻咽癌细胞克隆株差异表达cDNA消减文库,从随机测序10个克隆中获得6个在低转移鼻咽癌细胞株中高表达cDNA序列,它们都与已知的人类基因片段具有高度同源性。结论:应用抑制消减杂交技术,从一对具有高低转移表型差异的鼻咽癌细胞株中获得6条可能与肿瘤转移抑制相关的cDNA序列,它们可能在维持肿瘤细胞的稳定和抑制肿瘤转移中起重要作用。

Objective： To clone and identify metastasis-associated genes in nasopharyngeal carcinoma （NPC） and understand the molecular mechanisms underlying the metastasis of nasopharyngeal carcinoma. Methods： The suppression subtractive hybridization （SSH）. T/A cloning technique. DNA sequencing and bioinformatics were employed in this study, and two NPC cell strains were used as the study subject. CNE-2Z-H5-9 strain with high metastatic potential and CNE-2Z-H5 strain with low metastatic potential. Results： A different expression cDNA subtractive bank was successfully established from the human nasopharyngeal carcinoma cell line CNE-2Z; Six cDNA clones were highly expressed in the lower metastatic strain CNE-2Z-H5; aligned to sequences of the GenBank database by BLASTN. the six cDNA fragments were found to homologize highly with known human genes. Concluslon： Using SSH, we cloned and identified six cDNA fragments highly expressed in the lower metastatic nasopharyngeal carcinoma strain, and the genes represented by the six cDNA fragments might play a role in the metastatic process of nasopharyngeal carcinoma.