MAGE-1 Gene Expression Research Articles

Detection of tumor cells in peripheral blood of patients with gastric cancer using mRNA of MAGE genes as markers

To develop a sensitive and specific RT-PCR assay using the mRNA of MAGE-1 and MAGE-3 genes as specific tumor markers for the detection of the tumor cells in the peripheral blood of patients with gastric cancer. Peripheral blood was obtained from 40 patients with gastric cancer and from 20 healthy volunteers. The mRNA of MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMC) was detected by RT-PCR. The expressions of MAGE-1 and MAGE-3 mRNA in the tumor tissues of these gastric cancer patients were also detected by RT-PCR. Meanwhile,CEA expression by nested RT-PCR in PBMC of 40 gastric cancer patients was also detected. Of 40 gastric cancer patients, MAGE-1 and MAGE-3 mRNA were positive in 47.5% (19/40) and 25% (10/40) of PBMC respectively, and in 62.5% (25/40) and 30% (12/40) of gastric cancer tissues respectively. As a whole, in the PBMC of 40 gastric cancer patients, 25 (62.5%) samples were found to express at least one type of MAGE mRNA. In the patients whose tumors did not express MAGE-1 and/or MAGE-3 genes, the corresponding MAGE mRNA was also undetected in their PBMC. There was no expression of MAGE-1 or MAGE-3 gene in the PBMC from the 20 healthy donors. The positive rate of MAGE mRNA in PBMC was closely correlated with the tumor stage and lymph node metastasis (P <0.05). Positive rate of CEA gene expression was 32.5% (13/40) in the PBMC of 40 gastric cancer patients, 29 (72.5%)samples were detected to express at least one type of MAGE gene and CEA gene mRNA. MAGE-1, MAGE-3 and CEA mRNA are specifically detected with high percentage in the PBMC of gastric cancer patients by RT-PCR. They could be used as specific tumor markers for the detection of the circulating gastric cancer cells, and the detection results may be helpful to evaluate the prognosis of gastric cancer patients.

Genetically modified dendritic cells induced specific cytotoxity against human HCC cells in vitro

Objective: to transduce the tumor associated antigen gene MAGE-1 and/or IL-12 gene into dendritic cells (DC) and to observe the in vitro cytotoxic effect induced by the genetically modified DC against the human hepatocellular carcinoma (HCC) cell line SMMC7721. Methods: the MAGE-1 gene was inserted into the retrovirus vector LXSN to construct the recombinant retrovirus LMSN. The monocyte-derived DCs were transfected at appropriate differentiation stage by LMSN and/or a recombinant adenovirus AdmiL-12, which containing murine IL-12 gene. The control groups included retrovirus LXSN transfected, adenovirus AdBGFP transfected and non-transfected DCs. The MAGE-1 gene expression was identified by western blot and the mIL-12 p70 secretion was detected by ELISA assay. The in vitro cytotoxicities against SMMC7721 induced by genetically modified and control groups of DC were tested by MTT assay. Results: The MAGE-1 expression was detected by a monoclonal antibody in DCs tranfected with LMSN but not in control groups. At 16 h, 24 h and 48 h after transfection with AdmIL-12, the concentration of the mIL-12 p70 in the culture medium was 580pg/106 cells, 960pg/106 cells and 1100pg/106 cells respectively. The mIL-12 p70 secretions were not detected in other groups. The lytic activity (as judged by % lysis) induced by each groups of DC was 94.2±5.2% (LMSN and AdmIL-12 cotransfected group), 78.9±3.6% (LMSN transfected groups), 52.6±9.7% (AdmIL-12 transfected group), 34.7±4.3% (LXSN transfected group), 36.3±3.8% (AdBGFP transfected group) and 3.9±2.0% (non-transfected group) respectively. Except for LXSN transfected and AdBGFP transfected group, the difference of the lytic activities between other groups were statistically significant (P<0.05). Conclusion: The MAGE-1 gene modified DCs can induce relatively specific cytotoxicty against SMMC7721 in vitro and thus suggested that those genetically engineered DCs have the potential to serve as novel vaccine for HCC. Transduction of exogenous IL-12 gene into DCs may further enhance DCs’ activity and the effectiveness of the above DC vaccine.

Expression of melanoma antigen-encoding gene-1 predicts lymph node involvement in early gastric carcinomas.

Our previous study suggested that melanoma antigen-encoding gene-1 may be activated in gastric carcinomas when the tumors develop or invade. In the present work we studied the gene as a preoperative indicator of lymph node involvement in early gastric carcinoma. We used a reverse transcription-polymerase chain reaction to analyze expression of the gene in 47 endoscopic biopsy specimens from early gastric carcinomas. Relationships between gene expression and histopathologic findings were analyzed statistically. Eight of 47 tumor samples (17.0%) expressed the gene. The gene was not expressed in adjacent nontumor samples from carcinoma patients or in tissues from patients with benign gastric diseases. Gene expression correlated with infiltrative tumor growth in contrast to tumors with expansive growth patterns (P = 0.018). Gene expression also correlated with expression of platelet-derived growth factor-A (P < 0.001). MAGE-1 gene expression in biopsy specimens was a preoperative indicator of nodal involvement (P = 0.013). In conclusion, melanoma antigen-encoding gene-1 expression in biopsy specimens from early gastric carcinoma may serve preoperatively as an indicator of lymph node involvement.

MAGE, BAGE and GAGE: tumour antigen expression in benign and malignant ovarian tissue.

To determine if ovarian cancer patients would be suitable for MAGE-peptide vaccine-based immunotherapy, the frequency of expression of the MAGE-1-4 genes in ovarian tumours was assessed using reverse transcription polymerase chain reaction (RT-PCR) and product verification with digoxigenin-labelled oligonucleotide probes specific for each MAGE gene. In addition, the frequency of expression of more recently discovered tumour antigens (BAGE, GAGE -1, -2 and GAGE -3, -6) was established using RT-PCR and ethidium bromide staining. In this study 1/16 normal ovarian tissue specimens and 11/25 benign lesions expressed MAGE-1. In non-malignant tissue there was preferential expression of MAGE-1 in premenopausal women. A total of 15/27 malignant specimens expressed MAGE-1, including 10/14 serous cystadenocarcinomas. Expression of other tumour antigens was infrequent. The finding of MAGE-1 expression in both benign and malignant tissue questions previous assumptions regarding the role of MAGE genes in carcinogenesis. In addition, preferential MAGE-1 gene expression in non-malignant premenopausal tissue suggests that the MAGE genes may be involved in cellular proliferation as opposed to carcinogenesis or possibly that MAGE gene expression is under cyclical hormonal control. Finally, this study indicates that serous cystadenocarcinomas may be suitable tumours for MAGE-1 peptide immunotherapy.

Melanoma antigen-encoding gene-1 expression in invasive gastric carcinoma: correlation with stage of disease.

A human melanoma antigen-encoding gene-1, MAGE-1 gene, may be linked to the neoplastic transformation. In the present study, we extended this association with human gastric carcinomas. Specifically, we focused on the relationship between MAGE-1 gene expression and the histologic stage of gastric carcinoma. We used a reverse transcription-polymerase chain reaction assay (RT-PCR) to analyze the expression of the MAGE-1 gene in 38 endoscopic biopsy specimens from gastric carcinomas. We also studied the relationship between the expression of MAGE-1 gene and the genetic expression of several tumor invasion-related factors, including 72 kD type IV collagenase (MMP2), urokinase-type plasminogen activator (uPA), platelet-derived growth factor A (PDGF-A), and vascular endothelial growth factor (VEGF). Eleven of the 38 tumor samples (28.9%) expressed the MAGE-1 gene. MAGE-1 gene expression was present only in two of the 38 adjacent nontumor samples (5.3%). MAGE-1 gene expression in the 38 tumor samples was significantly correlated with the histological stage of disease (P = 0.0008), especially with the depth of histologically confirmed tumor invasion (t1 vs. t2 or greater, P = 0.00048). The expression of MAGE-1 gene correlated with the expression of MMP2 (P = 0.0064), uPA (P = 0.0390), and PDGF-A (P = 0.00018). These data suggest that the MAGE-1 gene may be activated in gastric carcinomas during periods of their development or invasion. In addition, a relationship between MAGE-1 gene expression and expression of invasion-related factors has been demonstrated.

High frequency of the MAGE-1 gene expression in hepatocellular carcinoma

The MAGE-1 gene, originally isolated from a human melanoma cell line, directs the expression of a potential tumor-rejection antigen, MZ2-E. This antigen is recognized by autologous cytotoxic T lymphocytes in association with a major histocompatibility complex class I molecule (HLA-A1), and has provided a basis for specific immunotherapy for melanoma patients. Here we show a high frequency of expression of the MAGE-1 gene in hepatocellular carcinoma (HCC). We examined the expression of the MAGE-1 gene in cell lines originated from hepatoma cells and in tumor and nontumor tissues of livers with HCC by reverse-transcription polymerase chain reaction (RT-PCR) and subsequent Southern blotting. MAGE-1 messenger RNA (mRNA) was expressed in three of four HCC cell lines (75%) and in 16 of 20 (80%) resected HCCs though none was detected in nontumor tissues. The high frequency expression of MAGE-1 gene in HCCs suggests the possibility as a target for tumor- specific immunotherapy for HCC patients. (Hepatology 1996 Dec;24(6):1437-40)

Differential MAGE-1 Gene Expression in Two Variants of an Erythroleukemic Cell Line (K562)

The MAGE-1 gene encodes an antigen recognized on melanoma cells by autologous cytotoxic T cells. This gene shows a wide range of expression in many human tumors but not in normal tissues except for testes. We used reverse transcription polymerase chain reaction assays to analyze the expression of the MAGE-1 gene in two variants of an erythroleukemic cell line, K562. Comparison of two variants of the K562 cell line in different stages of differentiation showed different patterns of expression of the MAGE-1 gene. The more undifferentiated cell line (K562A) expressed high levels of specific MAGE-1 mRNA, in contrast to K562B, which features of erythroid differentiation, without MAGE-1 expression. Interestingly, we could not modulate MAGE-1 gene expression when in vitro differentiation of K562A was induced with Ara-C. Finally, our data indicate that MAGE-1 expression is not necessary for the maintenance of the transformed phenotype.

P-486 Expression of MAGE-1 gene in hepatocellular carcinoma

P-486 Expression of MAGE-1 gene in hepatocellular carcinoma