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目的:应用SEREX技术筛选与鉴定鼻咽癌相关抗原,为鼻咽癌的早期诊断、免疫治疗和预后监测提供候选的生物学指标和理论基础。方法:从鼻咽癌新鲜活检组织中提取mRNA,构建cDNA表达文库。用鼻咽癌患者的血清筛选所构建的cDNA表达文库,获得的阳性克隆经扩增后提取噬菌粒DNA进行测序。所得序列与GenBank数据库中已知基因进行同源比较,分析其生物学特性。结果:所构建的cDNA原始文库含1×106个重组体,蓝白斑试验显示重组率为93%。用鼻咽癌血清对表达文库进行筛选,共获得21个阳性克隆,分别代表14个不同的cDNA插入片段。12个插入片段的DNA序列分别与基因库上的基因有不同程度的同源性,其中6个基因与鼻咽癌或其它肿瘤的发生、发展关系密切;另6个基因的生理功能未见文献报道。2个插入片段未发现有同源基因,推测可能为新发现的基因。结论:SEREX技术是目前筛选肿瘤抗原最简便、有效的手段之一。本文研究发现的抗原将为鼻咽癌的研究提供进一步的理论基础。
OBJECTIVE: To screen and identify nasopharyngeal carcinoma-associated antigens using SEREX technique to provide biological indicators and theoretical basis for the early diagnosis, immunotherapy and prognosis monitoring of nasopharyngeal carcinoma. Methods: mRNA was extracted from fresh biopsy tissues of nasopharyngeal carcinoma to construct cDNA expression library. The constructed cDNA expression library was screened by the serum of patients with nasopharyngeal carcinoma. The positive clones obtained were amplified and the phagemid DNA was extracted for sequencing. The obtained sequences were compared with the known genes in GenBank database to analyze their biological characteristics. Results: The original cDNA library contained 1 × 106 recombinants, and the blue-white test showed that the recombination rate was 93%. A total of 21 positive clones were obtained by screening the expression library with nasopharyngeal carcinoma serum, representing 14 different cDNA inserts respectively. The DNA sequences of the 12 insertions have different degrees of homology with the genes in the gene pool, of which 6 genes are closely related to the occurrence and development of nasopharyngeal carcinoma or other tumors. The physiological functions of the other 6 genes have not been reported Reported. Two inserted fragments found no homologous genes, speculated that may be newly discovered genes. Conclusion: SEREX technique is one of the most convenient and effective methods for screening tumor antigens. The antigens found in this study will provide further theoretical basis for the study of nasopharyngeal carcinoma.