Evaluation of MAGE-1 and MAGE-3 as tumour-specific markers to detect blood dissemination of hepatocellular carcinoma cells

Abstract

The members of MAGE gene family are highly expressed in human hepatocellular carcinoma (HCC). In the present study, we tested the tumour-specific MAGE-1 and MAGE-3 transcripts in the peripheral blood of HCC patients by nested RT -PCR to detect the circulating tumour cells and evaluate their potential clinical implication. Of 30 HCC patients, the positive rate of MAGE-1 and MAGE-3 transcripts was 43.3% (13 out of 30) and 33.3% (10 out of 30) in PBMC samples, whilst the positive rate was 70% (21 out of 30) and 53.3% (16 out of 30) in the resected HCC tissue samples, respectively. The positivity for at least one MAGE gene transcript was 63.3% (19 out of 30) in PBMC samples of HCC patients and 83.3% (25 out of 30) in the resected HCC tissue samples. MAGE-1 and/or MAGE-3 mRNA were not detected in the PBMC of those patients from whom the resected HCC tissues were MAGE-1 or MAGE-3 mRNA negative, nor in the 25 PBMC samples from healthy donors. The detection of MAGE transcripts in PBMC was correlated with the advanced stages and tumour size of the HCC, being 82.4% (14 out of 17) in tumour stages III and IVa, 56.6% (five out of nine) in stage II, and null (nought out of four) in stage I. The serum a-FP in 33.3% (10 out of 30) of HCC patients was normal or slightly elevated (540 ng ml 71). However, six of these 10 patients (a-FP 540 ng ml 71 ) were MAGE-1 and /or MAGE-3 mRNA positive in their PBMC. The follow-up survey of MAGE mRNA in PBMC was performed in 12 patients. Seven patients with persistent MAGE-1 and/or MAGE-3 mRNA positive or from negative turned to positive died because of metastasis and/or recurrence. In striking contrast, all four patients with MAGE-1 and/or MAGE-3 mRNA from positive turned to negative and one patient with persistent MAGE-3 transcript negative are alive after last test. Collectively, detection of MAGE transcripts with follow-up survey in PBMC is a feasible and reliable assay for the early prediction of the relapse and prognosis of the HCC patients.

Immunohistochemical analysis of the expression of FATE/BJ-HCC-2 antigen in normal and malignant tissues

“…Our previous reports also indicated that blood dissemination of tumor cells occurred in the early stage of HCC. 35 Detection of the FATE/BJ-HCC-2 gene transcript in PBMCs by nested PCR may be useful for specifically diagnosing micrometastasis of HCC cells in peripheral blood and benefit the prediction of HCC recurrence and prognosis of the disease after surgical treatment.…”

“…Early detection of metastatic tumor cells is critical to identify HCC patients at high risk of relapse and for the prescriptive therapy. 35 Since Smith et al 36 first successfully adopted the RT-PCR technique to assess tyrosinase mRNA as a tumor marker to detect circulating melanoma cells, the gene transcripts of both tissueassociated and tumor-specific markers have been applied in RT-PCR-based diagnosis of micrometastasis of tumor cells in peripheral blood, such as the tyrosinase and MAGE-3 transcripts in melanoma, MAGE-1 and MAGE-3 in HCC, etc. [35][36][37][38][39] The results in this study showed that the FATE/BJ-HCC-2 gene transcript was positive in PBMCs of 46.67% patients whose resected HCC tumor tissues were positive for FATE/BJ-HCC-2, implying the early dissemination of HCC cells in blood circulation.…”

Methylation profiling of twenty four genes and the concordant methylation behaviours of nineteen genes that may contribute to hepatocellular carcinogenesis

“…As shown the Fig 1 and Tab 2, although some samples were homozygously methylated or unmethylated, the majority had the co-existed methylated and unmethylated alleles. Inclusion of the normal liver tissues in our study enabled us to establish the normal methylation pattern of all the forty four genes, that except for the MAGEA1 [31] being homozygously methylated, the rest forty three were homozygously demethylated [27] (Fig 1 and Tab 2). Therefore, any changes in the methylation pattern from the normal liver tissue's to the non-cancerous neighboring tissues or HCC tissues should indicate the involvement of the DNA methylation mediated events during the HCC carcinogenesis, that have been scored as the positive methylation change.…”

“…Therefore, we have also assessed whether the switching-on of the otherwise hypermethylated genes in the normal cells is linked to the demethylated state of the promoter CpG island in the HCC tissues. The MAGEA1 gene is such an example, expression of which is off in the normal hepatocytes and on in HCC [31]. Correlating well with such an expression profile, we in deed found that the promoter CpG island of the MAGEA1 (panel 13, Fig 1) was fully methylated in the normal liver tissue, became unmethylated at a similar frequency in 18/28 (64%) and 21/28 (75%) in each of the paired HCC tissue samples, respectively.…”

Serum tumor markers for detection of hepatocellular carcinoma

“…It has been reported that the simultaneous determination of p53 antigen and anti-p53 antibodies has a sensitivity of 41.1% in the diagnosis of HCC [52] , and the over-expression of p53 in the serum or liver tissues of HCC patients prefigures the poorer prognosis and a shorter survival time (P = 0.0014) [52][53][54][55][56] . HCC patients with positive MAGE-1 or MAGE-3 mRNA die earlier because of metastasis or recurrence [57] . The sensitivity and specificity of serum human cervical cancer oncogene (HCCR) at the cut-off value of 15 µg/mL in detecting HCC are 78.2% and 95.7%, respectively.…”