Expression of MAGE-A9 in colorectal cancer and its clinical significance / 重庆医学

Objective To observe the expression change of melanoma-associated antigen(MAGE)-A9 in colorectal cancer (CRC)tissue and to explore its significance.Methods The samples in 23 cases of initially diagnosed CRC in the Affiliated Hospital of Nantong University from January 2006 to December 2008 were collected.The quantitative real-time(qRT)-PCR was adopted to detect MAGE-A9 mRNA expression in cancer tissue and corresponding paracancerous tissue.Its correlation with the clinicopatho-logical features and prognosis was analyzed.Results The positive rate of MAGE-A9 in CRC tissue was significantly higher than that in paracancerous normal tissue(P<0.05).MAGE-A9 protein expression in CRC was related to the clinicopathological features such as tumor differentiation degree(P=0.011),TNM stage(P=0.003),tumor infiltration depth(P=0.001)and lymph node me-tastasis(P=0.003).The survival analysis showed that the expression of MAGE-A9 was closely related to the prognosis of CRC pa-tients.Conclusion MAGE-A9 expression is increased in CRC tissue,suggesting the poor prognosis.

Human Leukocyte Antigen-A Allele Distribution in Nasopharyngeal Carcinoma Patients Showing Anti-Melanoma-Associated Antigen A or Synovial Sarcoma X-2 T Cell Response in Blood / 中华医学杂志(英文版)

<p><b>Background</b>Development of innovative immunotherapy is imperative to improve the poor survival of the nasopharyngeal carcinoma (NPC) patients. In this study, we evaluated the T cell response to melanoma-associated antigen (MAGE)-A1, MAGE-A3, or synovial sarcoma X-2 (SSX-2) in the peripheral blood of treatment-naive NPC patients. The relationship of responses among the three proteins and the human leukocyte antigen (HLA)-A types were analyzed to provide evidence of designing novel therapy.</p><p><b>Methods</b>Sixty-one NPC patients admitted into the Tumor Hospital affiliated to the Xinjiang Medical University between March 2015 and July 2016 were enrolled. Mononuclear cells were isolated from the peripheral blood before any treatment. HLA-A alleles were typed with Sanger sequence-based typing technique. The T cell response to the MAGE-A1, MAGE-A3, or SSX-2 was evaluated with the Enzyme-Linked ImmunoSpot assay. Mann-Whitney U-test was used to compare the T cell responses from different groups. Spearman's rank correlation was used to analyze the relationship of T cell responses.</p><p><b>Results</b>HLA-A*02:01, A*02:07, and A*24:02 were the three most frequent alleles (18.9%, 12.3%, and 11.5%, respectively) among the 22 detected alleles. 31.1%, 19.7%, and 16.4% of the patients displayed MAGE-A1, MAGE-A3, or SSX-2-specific T cell response, respectively. The magnitudes of response to the three proteins were 32.5, 38.0, and 28.7 SFC/10 peripheral blood mononuclear cells, respectively. The T cell response against the three proteins correlated with each other to different extent. The percentage of A*02:01 and A*24:02 carriers were significantly higher in patients responding to any of the three proteins compared to the nonresponders.</p><p><b>Conclusion</b>MAGE-A1, MAGE-A3, or SSX-2-specific T cell responses were detectable in a subgroup of NPC patients, the frequency and magnitude of which were correlated.</p>

Melanoma antigen A3 sourced from laryngeal cancer was expressed in mouse melanoma cell model / 中国免疫学杂志

Objective:To construct tumor cell model by determination of the pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid expressing steadily in mouse melanoma B16 cells.Methods:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid being constructed from the melanoma-associated antigen A3 genes sourcing laryngocarcinoma in advance was translated into the mouse melanoma B16 cells under the mediation of lipofectamine,and the positive clones were detected with G418.The expression of enhanced green fluorescent protein( EGFP) and MAGE-A3 mRNA in positive clones were detected by fluorescence microscopy and fluorescence quantitative PCR ( qRT-PCR ) assay, respectively.Results:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected into B16 cells successfully, the green fluorescence of fusion protein expression was found, and MAGE-A3 mRNA transcription in B16 cells expressions were detected in positive clones.Conclusion:The pIRES2-EGFP/MAGE-A3 eukaryotic expression plasmid has been transfected effectively and expressed stably by liposome method in the B16 cells.The expression of MAGE-A3 tumor cell model has been successfully established,which provide data for the study of laryngocarcinoma immunotherapy.

Effects of MAGEA3 inhibition by shRNA on apoptosis in human hepatocellular cancer cells / 中国病理生理杂志

AIM: To investigate the inhibitory effect of vector-based RNA interference(RNAi) on the expression of melanoma associated antigen A3(MAGEA3) protein in hepatocellular carcinoma cells and on apotposis of hepatocellular carcinoma cells.METHODS: A vector for transcribing specific small hairpin RNA(shRNA) targeting MAGEA3 gene was constructed,introduced into hepatocellular carcinoma MEL-ED1 cells by Lipofectamine 2000.The MAGEA3 protein and mRNA expression levels of MEL-ED1 cells were detected by Western blotting and RT-PCR, respectively.The cell apoptosis was studied by DNA fragmentation,electron microscopy,TUNEL assay,and annexin V/PI staining.RESULTS: The vector of RNA interference was successfully constructed and MAGEA3 expression was descreased significantly in MEL-ED1 cells.After the shRNA expression vector was transfected into the MEL-ED1 cells,the expression of MAGEA3 gene was inhibited significantly(by 90%).DNA fragmentation,electron microscopy and TUNEL assay showed classic apoptosis characters in the MEL-ED1 cells transfected with pSilencer-MAGEA3 plasmid with an apoptosis rate of 21.41% ?1.98%,significantly higher than those in the negative control group transfected with pSilencer-neo and in the non-transfected group(both P