人肝癌组织和外周血中甲胎蛋白基因片段扩增及临床应用

Amplification and clinical application of α-Fetoprotein gene fragment in human hepatoma tissues and peripheral blood

目的 研究外周血肝癌发生相关甲胎蛋白 (AFP)基因在肝癌早期诊断和鉴别诊断中的价值。方法 :从肝癌组织和肝病患者外周血中制备总RNA ,经随机引物和逆转录酶作用合成cDNA ,再以逆转录巢式聚合酶链反应 (RT -PCR)扩增AFP基因 ,并分析其临床价值。结果 RT -巢式PCR扩增的AFP基因片段为 15 9bp ,与原设计一致 ;在 2 0份肝癌及其癌周和远癌组织中 ,AFP基因阳性率分别为 10 0 %、6 5 %和 0 % ;肝癌患者外周血AFP基因检出率为 5 3.7% ,明显高于肝硬化、急、慢性肝炎和肝外肿瘤组患者 (P <0 .0 1) ;在I、II和III期肝癌中AFP基因阳性率分别为 33.3%、2 6 .3%和 71.8% ;伴肝外转移者全数阳性 ;AFP <5 0ng/ml肝癌AFP基因阳性率为 5 0 % ,且与肝癌大小无关。结论 外周血AFP基因检测是肝癌诊断与鉴别诊断、肝外转移或术后复发的有用标志物。

Objective To analyze alpha-fetoprotein (AFP) gene occurrence in human hepatocellular carcinomas (HCC) and its clinical value of early diagnosis and differential diagnosis in HCC. Methods Total RNAs was extracted from human hepatoma tissues and peripheral blood cells and synthesized to cDNA through random primers and reverse-transcriptase,amplified by nested reverse transcription PCR. The AFP gene fractions from patients with liver disease or non-liver disease were amplified, and its clinical values were analyzed in presnet study. Results The fragment of AFP-mRNA amplified by RT-PCR was 159bp and corresponded with the designed one. The incidences of AFP gene fragment in 20 human cancerous, paracancerous and distal cancerous tissues were 100%,65% and 0%, respectively.The positive rate of AFPmRNA in peripheral blood from patients with HCC were 53.7% and significantly higher than that in liver cirrhosis, chronic hepatitis, acute hepatitis and extrahepatic tumor ( P <0.01). The incidence of the fragment was 33.3% in I stage, 26.3% in II stage and 71.8% in III stage of HCC patients respectively.The AFPmRNA was 100% in all HCC patients with extrahepatic metastasis and 50% in HCC patients with serum AFP level less than 50 ng/ml.No correlation was found between tumor size and AFP gene. Conclusion The present data suggest that detection of AFP gene in peripheral blood is a useful marker for HCC diagnosis and differential diagnosis,extrahepatic matastasis and recurrence after operation.