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目的由于ＰＣＲ技术的应用，血循环中癌细胞的检测近有很大进展．本文用ＲＴＰＣＲ检测肝癌（ＨＣＣ）和其他慢性肝病患者外周血中ＡＦＰｍＲＮＡ，藉以反映ＨＣＣ细胞的存在，并与其他血清标记物比较．方法ＨＣＣ患者２２例，肝硬变和慢性乙型肝炎患者各１０例，健康成人受试者（对照）５例．取患者和对照的外周血，分离单核细胞，提取总ＲＮＡ并作电泳鉴定，用合成的两对引物进行巢式ＲＴＰＣＲ扩增ＡＦＰｍＲＮＡ，同时分析血清ＡＦＰ和乙肝标记物．结果ＡＦＰｍＲＮＡ在１３例ＨＣＣ（５９１％），２例肝硬变（２００％）患者外周血中测到，其余标本均为阴性．ＡＦＰｍＲＮＡ阳性的１３例患者肿瘤均大于５ｃｍ，为晚期患者．在该１３例患者中仅有６例（４６１％）在血清中测到ＡＦＰ，但有１２例（９２３％）ＨＢｓＡｇ，抗ＨＢｅ，抗ＨＢｃ全阳性，而ＡＦＰｍＲＮＡ阴性的５例该３标记物全阴性．结论ＲＴＰＣＲ扩增ＡＦＰｍＲＮＡ是检测ＨＣＣ和肝硬变患者循环肝癌细胞的敏感方法．患者外周血中的ＡＦＰｍＲＮＡ有可能作为肿瘤转移和复发的标记对ＨＣＣ诊断、随访观察和疗效评定有较大临床意义．
Objective The detection of cancer cells in the blood circulation has progressed greatly due to the application of PCR technology. In this study, AFP mRNA in peripheral blood of patients with hepatocellular carcinoma (HCC) and other chronic liver diseases was detected by RT-PCR to reflect the presence of HCC cells and compared with other serum markers. Methods Twenty-two patients with HCC, 10 patients with liver cirrhosis and 10 patients with chronic hepatitis B, and 5 healthy adult subjects (control). Peripheral blood was taken from patients and controls, mononuclear cells were isolated, total RNA was extracted and electrophoresis was performed. AFP mRNA was amplified by nested RT-PCR using two pairs of synthesized primers, and serum AFP and hepatitis B markers were analyzed at the same time. Results AFP mRNA was detected in peripheral blood of 13 cases of HCC (59.1%) and 2 cases of cirrhosis (20.0%). The rest of the specimens were negative. The tumors of 13 patients with positive AFP mRNA were all greater than 5 cm and were advanced patients. Of the 13 patients, only 6 (46.1%) had AFP detected in the serum, but 12 (92. 3%) had HBsAg, anti-HBe, and anti-HBc were all positive, while 5 were negative for AFP mRNA. 3 markers are all negative. Conclusion RT-PCR amplification of AFP mRNA is a sensitive method for detecting circulating hepatoma cells in patients with HCC and cirrhosis. AFPmRNA in the peripheral blood of patients may be used as a marker of tumor metastasis and recurrence. It has great clinical significance for the diagnosis, follow-up observation and therapeutic evaluation of HCC.