正常和肿瘤组织中ZNF165 mRNA的表达及其自发性抗体检测

目的:分析正常和肿瘤组织中ZNF165的表达及其在肝癌患者体内诱导产生的自发性体液免疫反应.方法:采用RT-PCR和定量PCR方法检测16种正常和4种肿瘤(肝癌、胃癌、结肠癌、肺癌)组织中ZNF165 mRNA及表达丰度;将ZNF165基因在原核细胞中进行蛋白表达,亲和层析法纯化重组ZNF165蛋白.Western blot方法对ZNF165在82例肝癌患者体内诱导的自发性抗体进行检测.结果:正常组织中ZNF165 mRNA只在睾丸组织表达,肝癌、胃癌、结肠癌和肺癌组织中ZNF165 mRNA阳性率分别为52.4%、 42.8%、 42.8%和21.4%.定量 PCR结果显示ZNF165 mRNA以高水平在肝癌组织与睾丸组织中表达,而在配对的癌旁组织与其他正常组织中仅存在很低水平的表达(P均＜0.01).82例肝癌患者血清中4例与ZNF165蛋白出现免疫反应,阳性率4.9%;36例正常人血清均为阴性.结论:ZNF165是一个新的肿瘤-睾丸抗原基因,它在机体内可诱导体液免疫反应,有可能作为肿瘤免疫治疗的候选抗原.

Detection of ZNF165 mRNA expression in normal tissue and tumor tissue and its spontaneous antibody in sera

Aim:To study ZNF165 expression in normal and tumor tissues and spontaneous humoral immunoresponse elicited by ZNF165 in liver cancer patients.Methods:ZNF165 mRNA and its quantity were measured in normal and tumor tissue by RT-PCR and quantitative PCR.Recombinant ZNF165 protein was expressed in prokaryotic cells and purified with affinity chromatography.Spontaneous antibody induced by ZNF165 in sera of liver cancer patients was detected by Western blot.Results:In normal tissues,ZNF165 mRNA only expressed in testis.In liver,stomach,colon,and lung cancer tissues,the positive rates of ZNF165 mRNA were 52.4%,42.8%,42.8%,and 21.4%,respectively.The results of quantitive PCR showed that ZNF165 mRNA expression was richer in liver cancer and testis tissue than that in cancer-adjacent tissue and other normal tissue(P<0.01).Of 82 sera from liver cancer patients,4 were reactive with recombinant ZNF165 protein,and the positive rate was 4.9%.Of 36 sera from normal person,the positive rate was 0.Conclusion:ZNF165 is a new cancer-testis antigen,which could elicit specific humoral immunoreponse in patients.It may be candidate antigen for tumor immunotherapy.