逆转录巢式PCR扩增AFP基因及肝癌诊断  

Amplification of Peripheral Blood Mononuclear Cell AFP Gene by Reverse Transcription Nested-PCR in HCC Diagnosis

作　　者：董志珍[1] 姚登福[1] 杨大明[1] 朱云松[1] 蒋道荣[1] 陆建新[1] 

机构地区：[1]南通医学院附属医院,南通226001

出　　处：《中国肿瘤》2000年第6期284-286,共3页China Cancer

基　　金：本课题受江苏省科委项目资助!(BS98029)

摘　　要：［目的］探讨外周血甲胎蛋白（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（ＰＣＲ）扩增ＡＦＰ基因片段，并分析其在肝癌诊断中的临床 应用价值。［结果］所设计ＲＴ－巢式ＰＣＲ扩增ＡＦＰ基因片段为 １５９ｂｐ，方法的灵敏度为５ｐｇ／ｍｌ。ＡＦＰ基因在肝癌、癌周和远癌组织中 的检出率分别为１００％、６５％和０；在肝癌患者外周血中的阳性率为５３．７％，明显高于其它肝病组和肝外肿瘤组（Ｐ＜０．０１）；在Ｉ、 Ⅱ和Ⅲ期肝癌中阳性率分别为３３．３％、２６．３％和７１．８％，伴肝外转移肝癌中全数阳性，与肿瘤大小无明显相关性。［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。Purpose]To explore the clinical value of pheripheral blood alphafetoprotein (AFP) gene analysis in diagnosis of hepatocellular carcinoma(HCC ). [Methods]Total RNAs was extracted from human hepatoma tissues and peripheral blood mononuclear cells, synthesized to cDNA through random primers and rever setranscriptase .The gene fragment of AFP was amplified by a nested-PCR assay and clinical value of its application was investigated in patients with liver cancer. [Results] The final product of AFP-mRNA amplified by RT-PCR was 159bp and the sensitivity of the assay was 5pg/ml .The incidence of AFP gene fragment were 100% in cancerous tissues, 65% in paracancerous tissues and 0 in distal cancerous tissues, respectively .The incidence of AFP-mRNA in periphery blood was 53. 7% in HCC patients and was significantly higher (P < 0. 01 ) than that in patients with other liver diseases or extra-hepatic tumors .The incidence of AFP-mRNA in HCC patients was 33. 3% in I stage. 26. 3% in Ⅱ stage and 71. 8% in Ⅲ stage. respectively .The AFP-mRNA was found in 4 patients with extra-hepatic metastasis(100%) No correlation was found between tumor size and AFP-mRNA. [Conclusion]The Present data suggest that the analysis Of AFP-mRNA is a useful gene marker for HCC diagnosis and monitoring of HCC with distal inatastasis or postoperative recurrence.

关 键 词：肝癌 AFP基因 RT-巢式PCR 诊断 

分 类 号：R735.704[医药卫生—肿瘤]