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

Mou DC, Cai SL, Peng JR, Wang Y, Chen HS, Pang XW, Leng XS, Chen WF - Br. J. Cancer (2002)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Center of Hepatobiliary Surgery, People's Hospital, Peking University Health Science Center, 42 Beilishilu, Beijing 100044, China.

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 (nought out of four) in stage I. The serum alpha-FP in 33.3% (10 out of 30) of HCC patients was normal or slightly elevated (< 40 ng ml(-1)). However, six of these 10 patients (alpha-FP < 40 ng ml(-1)) 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.

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Electrophoresis of second round of PCR products amplified from cDNA of PBMC samples of HCC patients. (1) M: molecular marker, 100 bp DNA ladder (Gibco); (2) P: Mel-Ed1, Positive control of nested MAGE-1 (299 bp) and MAGE-3 (371 bp) transcripts; (3) B: Blank control, PCR amplification in the absence of template; (4) β-actin (613 bp), cDNA quality control, which was amplified for one round of 35 cycles and all the samples shown here were positive; (5) Patient No. 1, 2, 3, 4, 10 showed MAGE-1 transcript positive, while patient No. 1, 2, 3, 14, 16 showed MAGE-3 transcript positive: (6) The second band of MAGE-3 transcript shown in Patient No. 1 and 14 was the genomic DNA with the size of 452 bp.
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fig2: Electrophoresis of second round of PCR products amplified from cDNA of PBMC samples of HCC patients. (1) M: molecular marker, 100 bp DNA ladder (Gibco); (2) P: Mel-Ed1, Positive control of nested MAGE-1 (299 bp) and MAGE-3 (371 bp) transcripts; (3) B: Blank control, PCR amplification in the absence of template; (4) β-actin (613 bp), cDNA quality control, which was amplified for one round of 35 cycles and all the samples shown here were positive; (5) Patient No. 1, 2, 3, 4, 10 showed MAGE-1 transcript positive, while patient No. 1, 2, 3, 14, 16 showed MAGE-3 transcript positive: (6) The second band of MAGE-3 transcript shown in Patient No. 1 and 14 was the genomic DNA with the size of 452 bp.

Mentions: With two-round RT–PCR amplification, the positive rate in blood samples was 43.3% (13 out of 30) for MAGE-1 transcript and 33.3% (10 out of 30) for MAGE-3 transcript. 63.3% (19 out of 30) of the PBMC samples were positive for at least one type of the two MAGE gene transcripts (Table 1). Among 25 cases of MAGE-1 and/or MAGE-3 mRNA positive HCC tissue samples, at least one of the two MAGE transcripts was detected in the PBMC of 19 cases of patients. The correlation positive rate was 76% (19 out of 25) between two distinct sources of specimen. MAGE-1 or MAGE-3 transcript was not detected in the PBMC of the patients whose resected HCC tissue samples were negative for MAGE-1 or MAGE-3 mRNA. In control samples, MAGE-1 or MAGE-3 mRNA could not be detected in PBMC from 25 healthy donors. The MAGE-1 and MAGE-3 positive PCR products amplified from mRNA of the PBMC samples were cloned into competent bacteria and three clones from each were randomly picked up and sequenced. The nucleotide sequence of MAGE-1 or MAGE-3 cDNA fragments were identical to the database of the GenBank. It confirms that the RT–PCR products were truly MAGE-1 and MAGE-3 cDNA (data not shown). The typical electrophoresis of nested RT–PCR products amplified from cDNA of PBMC samples of some HCC patients is shown in Figure 2Figure 2


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

Mou DC, Cai SL, Peng JR, Wang Y, Chen HS, Pang XW, Leng XS, Chen WF - Br. J. Cancer (2002)

Electrophoresis of second round of PCR products amplified from cDNA of PBMC samples of HCC patients. (1) M: molecular marker, 100 bp DNA ladder (Gibco); (2) P: Mel-Ed1, Positive control of nested MAGE-1 (299 bp) and MAGE-3 (371 bp) transcripts; (3) B: Blank control, PCR amplification in the absence of template; (4) β-actin (613 bp), cDNA quality control, which was amplified for one round of 35 cycles and all the samples shown here were positive; (5) Patient No. 1, 2, 3, 4, 10 showed MAGE-1 transcript positive, while patient No. 1, 2, 3, 14, 16 showed MAGE-3 transcript positive: (6) The second band of MAGE-3 transcript shown in Patient No. 1 and 14 was the genomic DNA with the size of 452 bp.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2746529&req=5

fig2: Electrophoresis of second round of PCR products amplified from cDNA of PBMC samples of HCC patients. (1) M: molecular marker, 100 bp DNA ladder (Gibco); (2) P: Mel-Ed1, Positive control of nested MAGE-1 (299 bp) and MAGE-3 (371 bp) transcripts; (3) B: Blank control, PCR amplification in the absence of template; (4) β-actin (613 bp), cDNA quality control, which was amplified for one round of 35 cycles and all the samples shown here were positive; (5) Patient No. 1, 2, 3, 4, 10 showed MAGE-1 transcript positive, while patient No. 1, 2, 3, 14, 16 showed MAGE-3 transcript positive: (6) The second band of MAGE-3 transcript shown in Patient No. 1 and 14 was the genomic DNA with the size of 452 bp.
Mentions: With two-round RT–PCR amplification, the positive rate in blood samples was 43.3% (13 out of 30) for MAGE-1 transcript and 33.3% (10 out of 30) for MAGE-3 transcript. 63.3% (19 out of 30) of the PBMC samples were positive for at least one type of the two MAGE gene transcripts (Table 1). Among 25 cases of MAGE-1 and/or MAGE-3 mRNA positive HCC tissue samples, at least one of the two MAGE transcripts was detected in the PBMC of 19 cases of patients. The correlation positive rate was 76% (19 out of 25) between two distinct sources of specimen. MAGE-1 or MAGE-3 transcript was not detected in the PBMC of the patients whose resected HCC tissue samples were negative for MAGE-1 or MAGE-3 mRNA. In control samples, MAGE-1 or MAGE-3 mRNA could not be detected in PBMC from 25 healthy donors. The MAGE-1 and MAGE-3 positive PCR products amplified from mRNA of the PBMC samples were cloned into competent bacteria and three clones from each were randomly picked up and sequenced. The nucleotide sequence of MAGE-1 or MAGE-3 cDNA fragments were identical to the database of the GenBank. It confirms that the RT–PCR products were truly MAGE-1 and MAGE-3 cDNA (data not shown). The typical electrophoresis of nested RT–PCR products amplified from cDNA of PBMC samples of some HCC patients is shown in Figure 2Figure 2

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Center of Hepatobiliary Surgery, People's Hospital, Peking University Health Science Center, 42 Beilishilu, Beijing 100044, China.

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 (nought out of four) in stage I. The serum alpha-FP in 33.3% (10 out of 30) of HCC patients was normal or slightly elevated (< 40 ng ml(-1)). However, six of these 10 patients (alpha-FP < 40 ng ml(-1)) 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.

Show MeSH
Related in: MedlinePlus