Hou Q, Chen K, Shan Z
Eur J Gynaecol Oncol. 2015;36(5):590-4.
To construct the cDNA library of the ascites tumor cells of ovarian cancer, which can be used to screen the related antigen for the early diagnosis of ovarian cancer and therapeutic targets of immune treatment.
Four cases of ovarian serous cystadenocarcinoma, two cases of ovarian mucinous cystadenocarcinoma, and two cases of ovarian endometrial carcinoma in patients with ascitic tumor cells which were used to construct the cDNA library. To screen the ovarian cancer antigen gene, evaluate the enzyme, and analyze nucleotide sequence, serological analysis of recombinant tumor cDNA expression libraries (SEREX) and suppression subtractive hybridization technique (SSH) techniques were utilized. The detection method of recombinant expression-based serological mini-arrays (SMARTA) was used to detect the ovarian cancer antigen and the positive reaction of 105 cases of ovarian cancer patients and 105 normal women's autoantibodies correspondingly in serum.
After two rounds of serologic screening and glycosides sequencing analysis, 59 candidates of ovarian cancer antigen gene fragments were finally identified, which corresponded to 50 genes. They were then divided into six categories: (1) the homologous genes which related to the known ovarian cancer genes, such as BARD 1 gene, etc; (2) the homologous genes which were associated with other tumors, such as TM4SFI gene, etc; (3) the genes which were expressed in a special organization, such as ILF3, FXR1 gene, etc; (4) the genes which were the same with some protein genes of special function, such as TIZ, ClD gene; (5) the homologous genes which possessed the same source with embryonic genes, such as PKHD1 gene, etc; (6) the remaining genes were the unknown genes without the homologous sequence in the gene pool, such as OV-189 genes.
SEREX technology combined with SSH method is an effective research strategy which can filter tumor antigen with high specific character; the corresponding autoantibodies of TM4SFl, ClD, TIZ, BARDI, FXRI, and OV-189 gene's recombinant antigen in serum can be regarded as the biomarkers which are used to diagnose ovarian cancer. The combination of multiple antigen detection can improve diagnostic efficiency.
构建卵巢癌腹水肿瘤细胞的cDNA文库，用于筛选卵巢癌早期诊断相关抗原及免疫治疗的靶点。
选取4例卵巢浆液性囊腺癌、2例卵巢黏液性囊腺癌及2例卵巢子宫内膜样癌患者的腹水肿瘤细胞构建cDNA文库。利用重组肿瘤cDNA表达文库的血清学分析（SEREX）和抑制性消减杂交技术（SSH）筛选卵巢癌抗原基因、评估酶活性并分析核苷酸序列。采用基于重组表达的血清学微阵列检测方法（SMARTA）检测卵巢癌抗原及105例卵巢癌患者和105例正常女性血清中相应的自身抗体阳性反应。
经过两轮血清学筛选及糖苷测序分析，最终鉴定出59个卵巢癌抗原基因片段候选物，对应50个基因。它们被分为六类：（1）与已知卵巢癌基因相关的同源基因，如BARD 1基因等；（2）与其他肿瘤相关的同源基因，如TM4SFI基因等；（3）在特定组织中表达的基因，如ILF3、FXR1基因等；（4）与某些具有特殊功能的蛋白质基因相同的基因，如TIZ、ClD基因；（5）与胚胎基因具有同源性的同源基因，如PKHD1基因等；（6）其余为基因库中无同源序列的未知基因，如OV - 189基因。
SEREX技术联合SSH方法是筛选高特异性肿瘤抗原的有效研究策略；血清中TM4SFl、ClD、TIZ、BARDI、FXRI及OV - 189基因重组抗原的相应自身抗体可作为诊断卵巢癌的生物标志物。多种抗原联合检测可提高诊断效率。
