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Cancer-testis (CT) genes are typically expressed in the testes; however, they have been linked to aberrant expression in a variety of malignancies. MAGE-B family genes are an example of CT genes. Therefore, the overarching objective of this study was to examine the expressions of MAGE-B family genes in several patients with colon cancer (CC) to see if they might be employed as cancer biomarkers in the early phases of cancer detection and to improve treatment. In this investigation, RT-PCR was used to analyze MAGE-B family genes in neighboring normal colon (NC) tissue from 10 CC patients. In addition, the effect of DNA demethylation on the expression status of the MAGE-B1 gene was evaluated by RT-PCR in HCT116 and Caco-2 cells and by qRT-PCR for HCT116 only after treating both CC cell lines with varying concentrations of 5-aza-2 ' -deoxycytidine (1.0, 5.0, and 10.0 μM) for 48 or 72 hours. All MAGE-B family genes except for MAGE-B1 showed weak bands in several samples of NC tissues: MAGE-B2, MAGE-B3, MAGE-B4, MAGE-B5, and MAGE-B6 genes were observed in 40%, 50%, 40%, 30%, and 60% of the NC samples, respectively. Nonetheless, they had strong bands in multiple samples of CC tissues, with 70%, 90%, 60%, 50%, and 90% of the CC samples, respectively. Interestingly, MAGE-B1 was detected in 60% of CC tissues but not in NC tissues, suggesting that it is a potential biomarker for early CC detection. MAGE-B1 expression was not observed in either untreated or DMSO-treated HCT116 cells after 48 or 72 hours of treatment. However, according to the RT-PCR and qRT-PCR results, the MAGE-B1 gene was overexpressed in the HCT116 cells treated with three different concentrations of 5-aza-2 ' -deoxycytidine. This shows that demethylation plays a crucial role in MAGE-B1 expression activation. [ABSTRACT FROM AUTHOR]

Colon cancer (CC) is a more frequent cancer type among men and is the third one among women. Cancer testis (CT) genes are classically expressed in the testes during postnatal life. However, they may be abnormally expressed in several types of cancers. The general purpose of this study was to look at the expressions of MAGE-A family genes, an example of the CT gene, in several patients with CC to evaluate whether they may be used as cancer biomarkers in the early stages of cancer identification and improve treatment options. In this study, MAGE-A family genes were examined using RT-PCR in 10 matching CC and normal colon (NC) samples. Also, the influence of DNA demethylation on the expression status of MAGE-A genes was assessed by treating the human CC cell lines with different doses of 5-aza-2′-deoxycytidine (1.0, 5.0, and 10.0 µM) for 48 or 72 h. All MAGE-A family gene results had faint bands in different samples of NC tissues, except for MAGE-A1. MAGE-A2 , MAGE-A3 , MAGE-A4 , MAGE-A6 , MAGE-A8 , MAGE-A9 , MAGE-A10 , MAGE-A11 , and MAGE-A12 genes were observed in 20 %, 60 %, 50 %, 80 %, 90 %, 60 %, 40 %, 30 %, and 40 % of the NC samples, respectively. However, they had strong bands in multiple samples of CC tissues with 80 %, 70 %, 60 %, 100 %, 100 %, 80 %, 60 %, 30 %, and 70 % of CC samples, respectively. Interestingly, MAGE-A1 was detected in 50 % of CC tissues but not in NC tissues, making this gene a suitable potential biomarker for early CC diagnosis. No expression of MAGE-A1 was detected in the Caco-2 cells after 48 h of treatment. However, the MAGE-A1 gene had faint bands after treatment of Caco-2 cells with 1–10 µM of 5-aza-2′-deoxycytidine for 72 h. In HCT116 cells, the MAGE-A1 gene had faint bands after 48 h and moderate-intensity bands 72 h after treating cells with 1–10 µM of -aza-2′-deoxycytidine. MAGE-A3 and MAGE-A4 were expressed after treating the Caco-2 cells with 1–10 µM 5-aza-2′-deoxycytidine for 48 or 72 h. Further protein-level tests and a larger cohort of patients are needed to assess this finding. [ABSTRACT FROM AUTHOR]