Detection of Hematogenous Micrometastasis in Hepatoma Patients by Nested Polymerase Chain Reaction

• Main Authors: Huang, Ming-Fa, 黃明發
• Other Authors: Yeh, Ming-Yang; Jiang, Shun-Yuan
• Format: Others
• Language: zh-TW
• Published: 1994
• Online Access: http://ndltd.ncl.edu.tw/handle/96567910960050541463

碩士 === 國防醫學院 === 微生物及免役學研究所 === 82 === Hepatocellular carcinoma (HCC) is the most common malignant solid tumor in Taiwan. It is one of the most difficult cancers to treat successfully. Tumor recurrence on the cirrhotic liver is frequent after surgical excision. Therefore, early diagnosis and elevate treatment efficiency is very importance. Alpha- fetoprotein (AFP) is a highly specific and sensitive marker for dignosis of HCC. The presence of tumor cells in the blood circulation may predict disease recurrence and metastasis. We have used nested polymerase chain reaction (nested-PCR) with primers specific for AFP gene to detect hepatoma cells in the blood. The AFP DNA fragment amplified from cDNA of Hep3 B hepatoma cells was used as a positive control. In a serial dilution experiment, nested-PCR could detect AFP mRNA from 1.9pg of Hep3B cDNA sample. In reconstitution experiments,AFP mRNA was detected in cDNA which prepared from one hepatoma cell per 25000 peripheral mononuclear cells. Peripheral mononuclear cells from sixteen hepatoma and seven hepatitis patients were further analyzed. Eight of the sixteen hepatoma patients and three of the seven hepatitis patients showed positive AFP mRNA expression. The positive AFP mRNA expression in the hepatoma patients was correlated with distant metastasis. The results indicated that nested-PCR is a sensitive method in detecting hematogenous hepatoma cells and may be valuable in predicting distant metastasis of the disease. In addition, the results showed that AFP mRNA was detected not only in hepatoma but also in hepatitis patients. Therefore, AFP is not a good target gene for the early diagnosis of hepatoma. Searching for hepatoma- specific target genes and combining the highly sensitive nested-PCR method for early diagnosis of hepatoma will become a research trend in the future.