从睾丸cDNA表达文库筛选胰腺癌肿瘤抗原

Search pancreatic cancer antigenes by screening a testis cDNA phage expression library with SEREX

目的利用胰腺癌患者血清筛选睾丸 cDNA 噬菌体表达文库,寻找胰腺癌特异性抗原,尤其是肿瘤-睾丸抗原。方法采用血清学筛选重组 cDNA 表达文库(SEREX)技术筛选睾丸cDNA 噬菌体表达文库。对获得的阳性克隆片段进行测序、鉴定和同源性比较。检测40例胰腺导管腺癌患者和40名健康对照血清中抗阳性克隆的抗体。结果筛选共获得107个阳性克隆,代表14个不同抗原基因,其中有13个基因与基因库中已知基因有很高的同源性,这些基因为 activin 受体AⅡ、LOC92912、KLHL12、IFI16和 CAGE 等。另有1个阳性克隆 HS1在基因库和 SEREX 数据库中均未发现有同源基因。胰腺癌患者组 HS1和 HS2克隆抗体阳性率显著高于健康对照组(分别15%与0%比较,X^2=4.50,P<0.05;22.5%与5%比较,X^2=5.16,P<0.05)。结论 HS1可能是1个新的肿瘤一睾丸抗原基因。有必要进一步研究这些阳性克隆的生物学功能和临床价值。

Objective To identify pancreatic cancer specific antigens, particularly cancer-testis antigens from a testis cDNA phage expression library by SEREX （ serological identification of antigen by recombinant cDNA expression libraries ）. Methods A testis cDNA phage expression library was screened. Positive clones were then seuenced and analysed. The antibodies to the positive Clones in sera from 40 pancreatic cancer patients and 40 healthy donors were assayed. Results 107 positive clones were obtained after immunoscreening. These clones were derived from 14 different genes. 13 genes were highly homologous to the genes known in GenBank. They are activin receptor A Ⅱ, LOC92912, KLHL12, IFI16, CAGE,and so on. One positive clone HS1 was found not to match any homological genes from GeneBank and SEREX databases. The prevalences of HS1 and HS2 antibodies in patients were significantly higher than those in healthy donors （ 15% vs 0% ,Χ^2 = 4.50, P 〈 0.05; 22.5% vs 5% ,Χ^2 = 5. 16, P〈0.05）. Conclusions HS1 might be a novel cancer-testis antigen gene. To explain biological function and clinical availability of these positive clones, further research is necessary.