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采用SEREX法筛选了自行构建的中国人卵巢癌cDNA表达文库 ,得到 2 7个阳性克隆 ,其中 3个为全长cDNA。序列分析和同源性比对结果表明所获得的 2 7个克隆分属于分化抗原、细胞结构蛋白及功能未知三类。选择其中一个全长cDNAMY OVA 13(该基因编码的蛋白是MAD2 ) ,利用基因工程方法克隆、表达和纯化得到目的蛋白 ,采用点杂交方法检测了MY OVA 13目的蛋白与正常人和肿瘤患者中的血清学反应。MY OVA 13抗体只在肿瘤组中检测到 (5 / 74 ) ,在正常组中均为阴性 ;间接ELISA测定结果显示 ,MY OVA 13抗体的水平在肿瘤组中均高于正常组 ,其中肝癌和前列腺癌组与正常组相比 ,在统计学上有显著差异 ,P值分别 <0 0 1和 <0 0 5。我们的初步结果表明MY OVA 13及其抗体具有一定的肿瘤特异性 ,有可能作为肿瘤诊断的标志物
SEREX cDNA was screened by self-constructed cDNA library of Chinese ovarian cancer, and 2 7 positive clones were obtained, of which 3 were full-length cDNA. Sequence analysis and homology comparison showed that 27 clones belonged to three types of differentiation antigen, cell structure protein and unknown function. One of the full-length cDNAMY OVA 13 (MAD2 encoding protein) was selected and cloned, expressed and purified by genetic engineering, and the dot blot method was used to detect the expression of MY OVA 13 protein in normal and cancer patients Serological reaction. The MY OVA 13 antibody was detected only in the tumor group (5/74), which was negative in the normal group. The indirect ELISA assay showed that the MY OVA 13 antibody level in the tumor group was higher than that in the normal group, and the liver cancer and There was a statistically significant difference between the prostate cancer group and the normal group, P <0 01 and <0 05, respectively. Our preliminary results indicate that MY OVA 13 and its antibodies have a tumor-specific potential that may serve as a marker of tumor diagnosis