Jiang S Y, Shyu R Y, Huang M F, Tang H S, Young T H, Roffler S R, Chiou Y S, Yeh M Y
Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China.
Br J Cancer. 1997;75(6):928-33. doi: 10.1038/bjc.1997.163.
The presence of tumour cells in the blood circulation may predict disease recurrence and metastasis. We have evaluated the specificity and sensitivity of detecting hepatoma cells in blood using nested polymerase chain reaction with primers specific for the alphafetoprotein (AFP) gene. The nested polymerase chain reaction amplified a 270-base pair AFP DNA fragment from cDNA of Hep 3B hepatoma cells. In a reconstitution experiment, AFP mRNA was detected from peripheral mononuclear cells isolated from 10 ml of blood containing as few as ten Hep 3B cells. Peripheral mononuclear cells from the blood of 20 hepatoma patients were analysed, and 19 patients showed positive AFP mRNA expression. Seven of 13 samples from hepatitis patients also showed positive AFP mRNA expression. All five paired samples of peripheral blood or umbilical cord blood from pregnant mothers and their babies, respectively, showed positive AFP expression. None of 22 control samples was positive. The presence of AFP mRNA in the blood of hepatitis or hepatoma patients suggests the presence of circulating hepatoma cells or hepatocytes in the circulation. The high incidence of AFP mRNA in the blood of hepatoma patients supports the notion of early haematogenous spreading of the disease.
血液循环中肿瘤细胞的存在可能预示疾病复发和转移。我们使用针对甲胎蛋白(AFP)基因的特异性引物,通过巢式聚合酶链反应评估了检测血液中肝癌细胞的特异性和敏感性。巢式聚合酶链反应从Hep 3B肝癌细胞的cDNA中扩增出一个270碱基对的AFP DNA片段。在一项重建实验中,从10毫升仅含10个Hep 3B细胞的血液中分离出的外周血单个核细胞中检测到了AFP mRNA。对20例肝癌患者血液中的外周血单个核细胞进行分析,19例患者显示AFP mRNA表达阳性。13例肝炎患者的样本中有7例也显示AFP mRNA表达阳性。分别来自孕妇及其婴儿的外周血或脐带血的所有5对配对样本均显示AFP表达阳性。22个对照样本均无阳性。肝炎或肝癌患者血液中AFP mRNA的存在提示循环中有循环肝癌细胞或肝细胞。肝癌患者血液中AFP mRNA的高发生率支持了该疾病早期血行播散的观点。