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目的筛选和分析肝细胞癌相关的肿瘤抗原。方法用肝细胞癌组织构建cDNA表达文库,通过重组cDNA表达文库血清学分析法,用自体患者和异体患者的血清对文库进行筛选。将所得阳性噬菌体克隆体内剪切获得其pBK-CMV噬菌粒。通过限制性核酸内切酶鉴定插入cDNA片段,并作测序和生物信息学分析。同时将阳性克隆中的LIMS1插入片段进行原核表达。结果自体血清筛选得到14种基因,异体筛选获得11种基因,两组中均筛选到kinectin,共有24种肝细胞癌相关的肿瘤抗原基因。其中有8种基因目前还不清楚其功能,其余16种基因根据确定的或者推断的功能可以分成8组。LIMS1原核重组蛋白表达成功。结论本研究为肝细胞癌的免疫治疗提供了候选基因,并为理解肝细胞癌发生和发展的机制提供了线索。
Objective To screen and analyze hepatocellular carcinoma-associated tumor antigens. Methods The cDNA expression library was constructed by using hepatocellular carcinoma (HCC) tissues. The library was screened by sera from autologous patients and allogeneic patients by serological analysis of recombinant cDNA expression library. The resulting positive phage clone was cloned in vivo to obtain its pBK-CMV phagemid. CDNA fragments were identified by restriction endonucleases and sequenced and bioinformatically analyzed. At the same time, positive clones were inserted into the LIMS1 fragment for prokaryotic expression. Results Fourteen genes were screened by autologous serum and eleven genes were screened by in vitro screening. Kinectin was screened in both groups, and 24 kinds of hepatocellular carcinoma-associated tumor antigen genes were found. Eight of these genes are not yet known for their function, and the remaining 16 genes are grouped into eight groups based on defined or inferred functions. LIMS1 prokaryotic recombinant protein expression was successful. Conclusion This study provides a candidate gene for the immunotherapy of hepatocellular carcinoma and provides clues for understanding the mechanism of the occurrence and development of hepatocellular carcinoma.