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目的利用肿瘤患者自体血清筛选患者肿瘤cDNA表达文库SEREX,寻找肿瘤特异性抗原是肿瘤免疫学研究的新策略,由此方法所获B细胞抗原肽可为进一步寻找T细胞抗原肽,开展肿瘤免疫治疗和诊断提供线索。方法采用SEREX方法,筛选中国人卵巢癌cDNA表达文库。对获得的阳性克隆片段测序鉴定和进行BLAST同源性比较。采用RTPCR,分析正常组织和肿瘤组织及肿瘤细胞株中6个新的EST片段的表达。结果所获基因片段大多数为已知抗原基因20/27。根据其来源又可归类为结构基因、线粒体基因和调控基因。另外,获得6个未知的EST片段。RTPCR的结果表明,4个ESTEST8788、1750、1754和3533片段在肿瘤和正常组织中均有表达,2个EST在正常组织中未见表达,而在肿瘤组织中有表达,可能是与肿瘤相关或特异的EST片段。结论SEREX方法不失为一种有效筛选卵巢癌肿瘤抗原的新方法,所获肿瘤抗原候选基因,可进一步进行结构和功能的研究。
Objective To screen the tumor cDNA expression library SEREX by autologous serum of tumor patients. The search for tumor-specific antigen is a new strategy for tumor immunology. The B cell antigen peptide obtained by this method can be used to further search for T cell antigen peptides and carry out tumor immunotherapy. And diagnoses provide clues. Methods The SEREX method was used to screen Chinese human ovarian cancer cDNA expression library. The obtained positive clone fragments were sequenced for identification and BLAST homology comparison was performed. RT-PCR was used to analyze the expression of six new EST fragments in normal tissues, tumor tissues and tumor cell lines. As a result, most of the gene fragments obtained were known antigen genes 20/27. According to their sources, they can be classified as structural genes, mitochondrial genes, and regulatory genes. In addition, 6 unknown EST fragments were obtained. RT-PCR results showed that the four ESTEST8788, 1750, 1754, and 3353 fragments were expressed in both tumors and normal tissues. The two ESTs were not expressed in normal tissues but were expressed in tumor tissues and may be related to tumors. Related or specific EST fragments. Conclusion The SEREX method is a new method for screening ovarian cancer tumor antigens. The tumor antigen candidate gene can be further studied for its structure and function.