肿瘤睾丸抗原CT10基因mRNA在肝细胞肝癌中的表达及HLA-A2限制性CTL表位的预测

Expression of cancer-testis antigen CT10 gene mRNA in hepatocellular carcinoma and prediction of HLA-A2 restricted CTL epitopes of CT10

目的检测CT10基因mRNA在肝细胞肝癌 (HCC)中的表达情况 ,了解肿瘤睾丸抗原CT10基因mRNA在HCC中的表达是否具有肿瘤特异性 ;对CT10HLA A2限制性CTL表位进行预测。方法用逆转录 聚合酶链反应 (RT PCR)方法对 45例HCC患者癌组织和相应癌旁组织的CT10基因mRNA表达情况进行了检测 ,对其中 3例RT PCR扩增产物的目的片段进行DNA序列测定 ;利用超基序法和量化基序法联合预测CT10HLA A2限制性CTL表位。结果 45例HCC患者中 ,19例表达CT10mRNA ,阳性率为 42 % ,相应的癌旁组织中结果均显示阴性 ;3例DNA测序结果表明RT PCR产物确为CT10cDNA。CT10的表达与年龄、性别、肿瘤体积、分化程度、血清甲胎球蛋白 (AFP)水平、HBV和HCV感染无显著相关性 (P >0 0 5 ) ;预测发现 9个HLA A2限制性CTL表位。结论CT10在HCC中呈高比例、特异性表达 ,可望成为HCC免疫治疗的理想靶位。

ObjectiveTo investigate the expression of cancer-testis antigen CT10 gene mRNA in hepatocellular carcinoma and predict the HLA-A2-restricted CTL epitopes of CT10.Methods The expression of CT10 mRNA was detected by using RT-PCR in HCC tissues and the corresponding adjacent non-HCC tissues from 45 HCC patients, among those 3 samples selected randomly from CT10 PCR positive products were sequenced.HLA-A2-restricted CTL epitopes of CT10 was predicted by peptide supermotif prediction combined with quantitative motif. Result CT10 mRNA was detectable in 19/45 (42.2%) of HCC samples, while the corresponding adjacent non-HCC tissue were all negative in expression of CT10 mRNA. In addition, The DNA sequence confirmed that the RT-PCR products were truly CT10 cDNA. No relationship was found between the expression of CT10 and demographic and clinical features such as age, sex, tumor size, degree of tumor differentiation, serum α-fetoprotein level and infection of hepatitis B virus or hepatitis C virus(P>0.05). Nine HLA-A2-restricted CTL epitopes of CT10 were found. Conclusion CT10 mRNA is expressed with a high percentage and specificity in hepatocellular carcinomas. The CT10 gene product is potential target for antigen-specific immunotherapy of hepatocellular carcinoma.