巢式RT-PCR检测肝癌患者外周血微量转移癌细胞  

Establishment of nested RT-PCR to detect circulating hepatocellular carcinoma cells in peripheral venous blood

作　　者：朱广迎[1] 陈杰[2] 朱斌[3] 蒋晓东[1] 

机构地区：[1]徐州医学院肿瘤研究所,江苏徐州221002 [2]徐州医学院影像系肿瘤基因实验室 [3]徐州医学院生物学教研室

出　　处：《徐州医学院学报》2001年第1期1-3,共3页Acta Academiae Medicinae Xuzhou

基　　金：江苏省卫生厅资助课题(Z9709)

摘　　要：目的　建立肝癌患者外周血微转移癌细胞的分子检测方法。方法　从肝癌患者外周血中分离有核细胞 ,用TRzol提取RNA ,特异引物扩增AFPmRNA ,凝胶电泳观察结果。分析AFPmRNA扩增结果与患者临床指标的关系。结果　肝癌患者外周血有核细胞RNA提取成功率为 10 0 %,33例肝癌患者AFPmRNA阳性率为 6 9%(2 3/ 33) ,临床肝内门静脉癌栓、肝外转移患者AFPmRNA扩增阳性率为 10 0 %,临床无转移患者AFPmRNA阳性率为 5 6 %(15 / 2 7) ,P <0 .0 5 ,AFPmRNA阳性患者的肿块最大径平均为 10 .6 7cm ,AFP >40 0 μg/L者占 40 %(8/ 2 0 ) ,AFPmRNA阴性患者肿块最大径平均为 7.13cm ,AFP >40 0 μg/L者占 10 %(1/ 10 )。 结论　巢式RTObjective To establish the molecular method to detect the circulating hepatocellular carcinoma (HCC) cells in peripheral venous blood. Methods Ficoll was used to isolate the mononuclear cells in peripheral venous blood of HCC patients. TRIzol was used to isolate the RNA. The specific primer pairs and nested RT-PCR were used to amplify the AFP mRNA. Results RNA was successfully isolated from all blood samples of the HCC patients. The positive rate of human AFP mRNA was found to be 69% (23/33). AFP mRNA was detected more commonly in patients with intra-portal venous carcinoma embolism or extra-hepatic metastasis (6/6,100%) than in those without metastasis (15/27,56%). The AFP mRNA positive patients had larger cancer masses and higher AFP values than the AFP mRNA negative patients. Conclusion Nested RT-PCR is a sensitive method to detect micrometastatic cancer cells in peripheral blood of HCC patients.

关 键 词：肝肿瘤 转移 巢式RT-PCR 

分 类 号：R735.7[医药卫生—肿瘤] R730.43[医药卫生—临床医学]