以MAGE基因mRNA为特异标记检测肝癌患者外周血肿瘤细胞

mRNA of MAGE genes as specific markers in detection of tumor cells in the peripheral blood of patients with hepatocellular carcinoma

目的 以黑色素瘤抗原 1(MAGE 1)和MAGE 3基因mRNA为特异性标记物 ,检测肝细胞癌 (HCC)患者外周血中的肿瘤细胞 ,并探讨其临床意义。 方法 采集 2 5例HCC患者及 2 0例健康志愿者的外周血 ,用巢式RT PCR方法检测外周血单核细胞 (PBMC)中MAGE 1和MAGE 3基因mRNA。同时用RT PCR方法检测HCC患者肝癌组织中上述MAGE基因的表达。 结果 4 4 % (11/2 5 )和 36 % (9/ 2 5 )HCC患者的PBMC中可分别检测到MAGE 1和MAGE 3基因mRNA ,而相应肝癌组织中MAGE 1和MAGE 3的表达率分别为 6 8% (17/ 2 5 )和 5 6 % (14 / 2 5 ) ;在 6 4 % (16 / 2 5 )的HCC患者PBMC中至少可检测到一种MAGE基因mRNA ;肝癌组织中不表达MAGE基因的病例以及 2 0例健康志愿者的PBMC中均未测出MAGE基因mRNA。HCC患者PBMC中两种MAGE基因mRNA的检出率与肿瘤TNM分期、直径密切相关 ,而与肿瘤分化程度、血清AFP水平无相关性。但 9例血清AFP正常或轻度升高者 (<5 0ng/ml) ,6例的PBMC中MAGE 1和 /或MAGE 3mRNA为阳性。 结论 MAGE 1和MAGE 3基因mRNA可作为特异性肿瘤标记物用于检测HCC患者PBMC中的肝癌细胞 ,巢式RT PCR的敏感性及特异性较高 ,其检测结果可能有助于判断HCC患者的预后。

Objective To detect tumor cells in the peripheral blood of patients with hepatocellular carcinoma (HCC) by using the mRNA of the MAGE 1 and MAGE 3 genes as specific tumor markers. Methods Peripheral blood was obtained from 25 HCC patients and 20 healthy volunteers. The mRNA of the MAGE 1 and MAGE 3 genes in the peripheral blood mononuclear cells (PBMCs) was detected by nested RT PCR. The MAGE 1 and MAGE 3 transcripts in the tumor tissues of these HCC patients were also detected by RT PCR. Results Of the 25 HCC patients , MAGE 1 and MAGE 3 mRNA were positive in 44% (11/25) and 36% (9/25) of PBMCs respectively, and in 68% (17/25) and 56% (14/25) of HCC tissues respectively. In the PBMCs of the 25 HCC patients, 16 (64%) samples were detected to express at least one type of MAGE mRNA. MAGE mRNA were not detected in the PBMCs from the patients whose tumors did not express the MAGE genes, nor in the PBMCs from the 20 healthy donors. The positive rate of MAGE mRNA in the PBMCs was closely correlated with the TNM stages and the diameter of tumors, but there was no correlation between the positive rate of MAGE mRNA in PBMCs and tumor differentiation degree or serum α FP level. Of 9 HCC patients whose serum α FP was normal or slightly elevated (<50 ng/ml), 6 were MAGE 1 and/or MAGE 3 mRNA positive in their PBMCs. Conclusion MAGE 1 and MAGE 3 mRNA could be specifically detected with high percentage in the PBMCs of HCC patients by our method. They can be used as specific tumor markers for the detection of the circulating HCC cells, and the detection results may be helpful to evaluate the prognosis of HCC patients.