Expression Profile of MAGE-B1 Gene and Its Hypomethylation Activation in Colon Cancer

Abstract

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.

HSPB6 Is Depleted in Colon Cancer Patients and Its Expression Is Induced by 5-aza-2′-Deoxycytidine In Vitro

“…We obtained two ethical approvals to collect samples from patients with CC and induce CC formation in male Wistar rats from the Al-Imam Muhammad Ibn Saud Islamic University (HAPO-01-R-011, July 9, 2020) [ 14 ] and the King Saud University Ethics of Scientific Research Committee (KSU-SE-20-20. May 8, 2020) [ 30 ], respectively.…”

“…May 8, 2020) [ 30 ], respectively. Ten patients with CC signed a consent form and provided us with their ages, personal medical information, and social behavior [ 14 ]. The 10 male Wistar rats were provided by the animal house at the College of Science, King Saud University [ 30 ].…”

“…Colon tissues from male Wistar rats were also taken from the normal control group and the DMH-induced group [ 31 ]. All tissues were immediately placed in Eppendorf tubes containing RNAlater [ 14 , 31 ].…”

“…Despite the ambiguity of the pathological pathways of CC initiation, some abnormal genes are implicated in accelerating the growth of CC and are involved in chemoresistance, including CXCL12 , SUL1A1 [ 9 ], ADAM2 , SPZ1 [ 10 ], CTAG1A , and TKTL2 [ 11 ]. Additionally, epigenetic events, particularly DNA methylation, contribute to CC tumorigenesis, either by silencing tumor suppressor genes SEPT9 [ 12 ] or FOXO1 [ 13 ] or activating oncogenes MAGE -B1 [ 14 ], MYBL2 , and AXIN2 [ 15 ]. Thus, investigating the alternation of gene expression and epigenetic events during tumorigenesis could provide promising targets that can be utilized as predictive, prognostic, and diagnostic biomarkers [ 16 ].…”

Higher Expression Levels of SSX1 and SSX2 in Patients with Colon Cancer: Regulated In Vitro by the Inhibition of Methylation and Histone Deacetylation

“…Colon cancer (CC) is the third and fourth most common cause of cancer-related death worldwide among males and females, respectively [ 1 ]. In Saudi Arabia, it is the leading cause of mortality in both sexes and ranks as the first and third most frequently diagnosed malignancy in men and women, respectively [ 2 ]. Furthermore, the prevalence rate of CC is high among Saudi men and women between the ages of 55 and 58 years [ 3 ].…”