联合免疫磁珠与逆转聚合酶联反应在肝细胞癌患者外周血癌细胞检测中的应用

Application of immunomagnetic bead sorting and nested reverse transcriptase-polymerase chain reaction for detection of the cancer cells in peripheral blood of patients with hepatocellular carcinoma .

目的研究联合免疫磁珠分类与巢式逆转录-聚合酶链反应(RT-PCR)在肝细胞癌患者外周血癌细胞检测中的应用价值。方法分别收集肝细胞癌(45例)、肝硬化(16例)、肝炎(20例)患者及健康人(18例)的外周血,以CD45和Ber-EP4免疫磁珠富集血液中的癌细胞,用巢式RT-PCR检测其外周血甲胎蛋白(AFP)mRNA水平。结果AFPmRNA在肝细胞癌、肝硬化和肝炎患者的检出率分别为72.1%、43.8%和25.0%,肝细胞癌组与肝硬化组和肝炎组比较的差异均有显著性意义(P<0.05和P<0.01)。结论联合免疫磁珠分类与巢式RT-PCR法可提高肝细胞癌患者外周血AFPmRNA的检出率。

Objective To study the applicantion of immunomagnetic bead sorting and nested reverse transcfiptase-polymerase chain reaction （ RT-PCR） for detection of the cancer cells in peripheral blood of patients with hepatocellular carcinoma（ HCC）. Methods Peripheral blood were collected from patients with HCC, liver cirrhosis （ LC ）, hepatitis and normal donors. The circulating cancer cells were enriched with CIM5 and BER-EP4 immunomagnetic beads. The alpha - fetoprotein（ AFP） mRNA was amplified with nested RT-PCR. Results The total postive rate of AFP mRNA was 72.1% ,43.8% and 25.0% in patients with HCC, LC and hepatitis, respectively. There was significant difference among the patients with HCC, LC and hepatitis （HCC vs LC, P〈0.05;HCC vs hepatitis,P〈0.01）. Conclusion Immunomagnetic bead sorting method and nested RT-PCR may raise the detective rate of AFP mRNA in peripheral blood of patients with HCC.