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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。
Objective To detect the expression of tumor testis antigen CT9 and CT10 mRNA in hepatocellular carcinoma (HCC). Methods The expression of CT9 and CT10 mRNA in HCC patients’ cancer tissues, adjacent paracancerous tissues and controls (cirrhosis and normal liver tissues) were detected by reverse transcription-polymerase chain reaction (RT-PCR) Four anti-HCV RT-PCR positive products were determined by DNA sequencing, and combined with AFP, anti-HCV, HBsAg, tumor diameter, differentiation and other clinical indicators. Results The positive rates of CT9 and CT10 mRNA in 56 HCC patients were 51.8% (29/56) and 44.6% (25/56), respectively. The positive rates of CT9 and CT10 mRNA expression were 66.1% (37 / 56) and 30.4% (17/56) respectively. The PCR products showed the same size as the positive control, while only CT9 mRNA expression was positive in adjacent tissues, the positive rate was 7.1% ( 4/56). However, the expression intensity was significantly weaker than that in the cancerous tissue. Further pathological examination showed no cancerous cells in adjacent tissues with CT9 positive expression. Tested 10 cases of cirrhosis and 10 cases of normal liver tissue were not detected CT9 and CT10 expression. DNA sequencing results show that the RT-PCR products indeed CT9 and CT10 cDNA. The expression of CT9 and CT10 had no significant correlation with age, sex, tumor size, differentiation, AFP level, HBV and HCV infection (P> 0.05). Whereas CT9 and CT10 genes are present in some AFP normal (<25 ng / L) HCC patients. Conclusions CT9 and CT10 genes are highly expressed and highly expressed in HCC, which provide a new ideal target for HCC immunotherapy. CT9 and CT10 are expressed in HCC at the same time. In order to apply multivalent tumor vaccine based on these tumor antigens HCC treatment provides an experimental basis.