外周血甲胎球蛋白mRNA扩增在肝癌诊断和鉴别诊断中的价值

Peripheral blood AFP mRNA amplification in the diagnosis and differential diagnosis of hepatocellular carcinoma

目的 探讨外周血甲胎球蛋白 (alpha fetoprotein ,AFP)基因分析对肝癌诊断的临床价值。方法 从人肝癌组织和外周血单核细胞中制备总RNA ,经随机引物和逆转录酶合成cDNA后 ,以巢式聚合酶链反应 (polymerasechainreaction ,PCR)扩增AFP基因片段 ,分析其在肝癌诊断中的临床应用价值。结果 所设计反转录巢式PCR扩增AFP基因片段为 15 9bp ,方法的灵敏度为 5 μg/L ;AFP基因在肝癌的癌灶组织、癌灶周围组织和远离癌灶肝组织中的检出率分别为 10 0 % (2 0 /2 0 )、6 5 %(13/2 0 )和 0 % (0 /2 0 ) ;在肝癌患者外周血中的阳性率为 5 4% (36 /6 7) ,明显高于其他肝病组和肝外肿瘤组 ;在Ⅰ、Ⅱ和Ⅲ期肝癌中阳性率分别为 33% (3/9)、2 6 % (5 /19)和 72 % (2 8/39) ;伴肝外转移肝癌 4例和术后复发肝癌 12例全部阳性 ;AFP基因与肝癌体积间无明显相关性。结论 分析外周血AFP mRNA有助于肝癌诊断、肝癌转移或术后复发的监测。

Objective[WT5”BZ] To explore the clinical value of pheripheral blood alpha-fetoprotein (AFP) gene analysis in the diagnosis of hepatocellular carcinoma (HCC). [WT5”HZ]Methods[WT5”BZ] Total RNAs were extracted from human hepatoma tissues and peripheral blood mononuclear cells, synthesized to cDNA through random primers and reverse transcriptase. The gene fragment of AFP was amplified by a nested-polymerase chain reaction (PCR) assay and clinical values of its application were investigated in patients with liver disease.[WT5”HZ] Results[WT5”BZ] The final products of AFP-mRNA amplified by reverse transcriptase (RT)-PCR were 159*!bp and the sensitivity of the assay was 5*!μg/L. The incidence of AFP gene fragment was 100% in cancerous tissues, 65% in paracancerous and 0% in hepatic tissues distal to the foci of cancer, respectively. The incidence of AFP-mRNA in peripheral blood was 53 7% in HCC patients, which was significantly higher than that in patients with other liver diseases or extra-hepatic tumors. The incidence of AFP-mRNA in HCC patients was 33 3% in stage Ⅰ, 26 3% in stage Ⅱ and 71 8% in stage[CM(422] Ⅲ, respectively. The incidence of AFP-mRNA was [CM(15]100% in 4 patients with extrahepatic metastasis and [CM(2]12 patients with postoperative recurrence. No correlation was found between tumor size and AFP-mRNA expression.[WT5”HZ] Conclusion[WT5”BZ] The present data suggest that the analysis of AFP-mRNA is a useful gene marker for HCC diagnosis and monitoring HCC with distal matastasis or postoperative recurrence. [WT5”HZ]