【摘要】 目的检测肿瘤睾丸抗原CT9和CT10基因mRNA在肝细胞癌(HCC)中的表达情况。方法用逆转录-聚合酶链反应(RT-PCR)方法对HCC病人的癌组织和相应癌旁组织及对照(肝硬化和正常肝组织)中CT9和CT10 mRNA的表达进行了检测,随机选取每一种抗原RT-PCR呈阳性的产物4例进行DNA序列测定,结合AFP、抗HCV、HBsAg、肿瘤直径、分化程度等临床指标进行分析。结果56例HCC病人中,CT9、CT10基因mRNA阳性率分别是51．8％(29／56)和44．6％(25／56),至少表达一种CTA者达66．1％(37／56),表达两种者为30．4％(17／56);电泳显示PCR产物与阳性对照大小一致;而癌旁组织中仅有CT9基因mRNA表达为阳性,阳性率为7．1％(4／56),但表达强度明显弱于在癌组织中的表达,进一步的病理检查未发现CT9表达呈阳性的癌旁组织中有癌细胞。所测10例肝硬化和10例正常肝组织均未检测到CT9和CT10的表达。DNA测序结果表明RT-PCR产物确为CT9和CT10 cDNA。CT9和CT10的表达与年龄、性别、肿瘤大小、分化程度、AFP水平、HBV和HCV感染无显著相关性(P>0．05)。而在部分AFP正常(<25 ng／L)HCC病人中存在CT9和CT10基因的表达。结论CT9和CT10基因在HCC中呈高比例、高特异表达,为HCC免疫治疗提供了新的理想靶位;CT9和CT10同时在HCC中表达,为应用以这些肿瘤抗原为基础的多价瘤苗治疗HCC提供了实验依据。更多还原

【Abstract】 Objective To investigate the expression of two cancer testis antigen (CT9 and CT10) gene mRNA in hepatocellular carcinoma (HCC) and explore the possibility of using CT9 and CT10 encoding protein as a target for immunotherapy in HCC patients. Methods The expression of CT9 and CT10 mRNA was detected using RT-PCR in HCC tissues and the adjacent non-HCC tissues from 56 HCC patients, liver samples from 10 patients with liver cirrhosis and 10 healthy individuals. Four samples selected randomly from CT9 and CT10 PCR positive products were sequenced. The relationship between positive expression rate of CT9 and cT10 gene and clinical and laboratory data including AFP, anti-HCV, HBsAg and the tumor diameter in HCC patients was also determined. Results CT9 and CT10 were detectable in 51. 8% (29/56) and 44. 6% (25/56) of HCC samples. Only weak expression of CT9 was found in 7.1% (4/56) of the adjacent non-HCC tissues and there was no metastatic lesion. Taking these 2 CT antigens together, at least one CT antigen gene mRNA was positive in 66. 1% (37/56) of HCC tissues, with synchronous expression of the 2 genes in 30. 4% (17/56) of HCC tissues. Meanwhile, the expression of CT9 and CT10 was not seen in the liver samples from 10 patients with liver cirrhosis and 10 healthy individuals. The DNA sequencing confirmed that the RT-PCR products were of true cDNA of CT9 and CT10. The expression of CT9 and CT10 was not related to clinical indicators such as age, sex, tumor size, tumor differentiation degree, serum AFP level and HBV or HCV infection (P>0. 05). However, specific expression of CT9 and CT10 was found in some patients with normal serum level of AFP (<25 ng/L). Conclusions CT9 and CT10 mRNA is expressed with a high percentage and specificity in HCC and their products are new potential promising targets for antigen-specific immunotherapy of HCC. Co-expression of CT9 and CT10 in HCC provides possibility of polyvalent vaccination for HCC.更多还原