摘要
目的：探讨卵巢癌相关肿瘤抗原新基因在卵巢癌诊断和免疫治疗中的应用价值。研究肿瘤抗原新基因与卵巢癌发生的关系及临床意义。
材料和方法：利用网上生物信息学分析软件对筛选到的180个卵巢癌肿瘤抗原基因克隆中的55个克隆的核苷酸序列进行分析，并检测其中8个新基因在卵巢癌患者和正常女性血清中的抗体。采用RT-PCR和原位杂交的方法研究卵巢癌相关肿瘤抗原新基因在卵巢癌组织和正常卵巢组织中的表达，分析其与卵巢癌临床指标的关系。
结果：55个克隆的核苷酸序列代表45个卵巢癌抗原基因，其中8个是国内外首次发现的在GeneBank中尚未登记的新基因，这些新基因都是具有免疫原性的功能基因。其中OCY-142新基因定位于2号染色体短臂13区(2P13)，是抑癌基因BARD1的可剪切变异体，OCY-142相应的蛋白质结构域比BRAD1少了NH_2-端的RING结构域和COOH-端BRCT结构域，可以被CD4~+和CD8~+T细胞识别。
8个卵巢癌肿瘤抗原新基因在卵巢癌患者和正常人血清中相应抗体的检测发现，其中7个在卵巢癌患者血清中的阳性率显著高于正常人，选择这7个卵巢癌肿瘤抗原新基因对卵巢癌患者和正常女性进行联合检测，其敏感性和特异性达到96.9％和95.2％。CA125异常的卵巢癌41例患者中，OCY-142单独检测的阳性率为95.12％，而CA125正常的卵巢癌57例患者中，OCY-142检
Objective: To evaluate the roles of novel antigen genes in diagnosis and immunothreapy of ovarian cancer and to analysis the relationship between novel antigen genes and the genesis of ovarian cancer.
Methods: The genes selected by SEREX and SSH in our previously research works were identified after analysis by bioinformatics. 8 novel antigen phage clones were screened using allogenic sera from 98 patients with ovarian serous papillary cystadenoma. We detected the expression of OCY-66, OCY-189 and OCY-142 in 28 ovarian cancers and 30 normal ovarian tissues by RT-PCR, and studied the expression of OCY-142 in these ovarian cancers and normal ovarian tissues by nonradioactive hybridization in situ.
Results: 55 positive clones were chosen randomly and sequenced. The sequencing results indicated that 55 positive clones represent 45 tumor antigen genes. 8 of them are novel genes, and they all have immunogenicity. OCY-142 is the novel-splicing variant of BARD1 (BRCA1 associated ring domain 1). It is located in chromosome 2 (2P13) and has one more intron sequences of BARD1 gene. It lost sequences coding for its RING domain and BRCT domain which are demonstrated responsible for interacting with the tumor suppressor gene BRCA1 and keeping its function. The immunogenicity of OCY-142 was also found derived from exposing of hide immuno-epitope by RING domain lost. OCY-142 may also be recognized by CD8~+T cell after MHC-I binding peptide prediction.
The expression positive ratio of 8 novel antigen phage clones presented from 53.1%~91.8%, but with very low positive ratio in sera of 98 health donors. Using a combination of novel antigen genes, the sensitivity and specificity for detecting ovarian cancer is 96.9% and 95.2%, respectively. 36.9% of ovarian cancers with normal CA125 level can be detected out by OCY-142.
引文
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