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

Sep 24, 2026

Colon cancer (CC) has high mortality and is frequently diagnosed at advanced stages in Saudi Arabia, making early detection biomarkers essential. This study evaluates cancer-testis (CT) genes from the SSX family by validating SSX1, SSX2, and SSX3 expression in CC tissues versus matched normal colons. RT-PCR and qRT-PCR assays analyzed 30 male Saudi samples and tested epigenetic regulation in vitro using 5-aza-20-deoxycytidine and trichostatin. SSX1 and SSX2 were absent in normal tissues but elevated in CC, while SSX3 was undetected. Treatment increased SSX gene expression in CC cells, suggesting SSX1 and SSX2 as candidate biomarkers and potential therapeutic targets.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages19

Reading Time48 min

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Abstract

Background and Objectives

Materials and Methods

Results

Conclusions

Introduction

medicina Article Higher Expression Levels of SSX1 and SSX2 in Patients with Colon Cancer: Regulated In Vitro by the Inhibition of Methylation and Histone Deacetylation Turki M. Alrubie 1, Abdullah M. Alamri 2, Bader O. Almutairi 1, Abdulwahed F. Alrefaei 1, Maha M. Arafah 3, Mohammad Alanazi 2, Abdelhabib Semlali 4 and Mikhlid H. Almutairi 1, * Citation: Alrubie, T.M.; Alamri, A.M.; Almutairi, B.O.; Alrefaei, A.F.; Arafah, M.M.; Alanazi, M.; Semlali, A.; Almutairi, M.H. Higher Expression Levels of SSX1 and SSX2 in Patients with Colon Cancer: Regulated In Vitro by the Inhibition of Methylation and Histone Deacetylation. Medicina 2023, 59, 988 . [https://doi.org/10.3390/](https://doi.org/10.3390/)[ ](https://doi.org/10.3390/)medicina59050988 Academic Editor: Vaidutis Kuˇcinskas Received: 26 April 2023 Revised: 13 May 2023 Accepted: 18 May 2023 Published: 20 May 2023 Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) . 1 Zoology Department, College of Science, King Saud University, Riyadh 11451, Saudi Arabia; [442106519@student.ksu.edu.sa](442106519@student.ksu.edu.sa) (T.M.A.); [bomotairi@ksu.edu.sa](bomotairi@ksu.edu.sa) (B.O.A.); [afrefaei@ksu.edu.sa](afrefaei@ksu.edu.sa) (A.F.A.) 2 Genome Research Chair, Department of Biochemistry, College of Science, King Saud University, Riyadh 11495, Saudi Arabia; [abdullah@ksu.edu.sa](abdullah@ksu.edu.sa) (A.M.A.); [msanazi@ksu.edu.sa](msanazi@ksu.edu.sa) (M.A.) 3 Pathology Department, College of Medicine, King Saud University, Riyadh 11451, Saudi Arabia; [marafah@ksu.edu.sa](marafah@ksu.edu.sa) 4 Groupe de Recherche en 􀂒cologie Buccale, Facult² de M²decine Dentaire, Universit² Laval, 2420 Rue de la Terrasse, Local 1758, Qu²bec, QC G1V 0A6, Canada; abdelhabib.semlali@greb.ulaval.ca * [Correspondence: malmutari@ksu.edu.sa](Correspondence: malmutari@ksu.edu.sa) Abstract: Background and Objectives: Colon cancer (CC) has a high mortality rate and is often diagnosed at an advanced stage in Saudi Arabia. Thus, the identiﬁcation and characterization of potential new cancer-speciﬁc biomarkers are imperative for improving the diagnosis of CC by detecting it at an early stage. Cancer-testis (CT) genes have been identiﬁed as potential biomarkers for the early diagnosis of various cancers. Among the CT genes are those belonging to the SSX family. In order to assess the usefulness of SSX family genes as cancer biomarkers for the detection of early-stage CC, the goal of this research was to validate the expressions of these genes in patients with CC and in matched patients with normal colons (NCs) . Materials and Methods: RT-PCR assays were used to analyze the SSX1, SSX2, and SSX3 family gene expression levels in 30 neighboring NC and CC tissue samples from male Saudi patients. Epigenetic alterations were also tested in vitro using qRT-PCR analysis to determine whether reduced DNA methyltransferase or histone deacetylation could stimulate SSX gene expression via 5-aza-20-deoxycytidine and trichostatin treatments, respectively. Results: The RT-PCR results showed SSX1 and SSX2 gene expression in 10% and 20% of the CC tissue specimens, respectively, but not in any of the NC tissue specimens. However, no SSX3 expression was detected in any of the examined CC or NC tissue samples. In addition, the qRT-PCR results showed signiﬁcantly higher SSX1 and SSX2 expression levels in the CC tissue samples than in the NC tissue samples. The 5-aza-20-deoxycytidine and trichostatin treatments signiﬁcantly induced the mRNA expression levels of the SSX1, SSX2, and SSX3 genes in the CC cells in vitro. Conclusions: These ﬁndings suggest that SSX1 and SSX2 are potentially suitable candidate

FAQs

Why are SSX family genes investigated in colon cancer?

Colon cancer is often detected at an advanced stage in Saudi Arabia, so early diagnosis biomarkers are needed. SSX family genes are cancer-testis (CT) genes that may be abnormally activated in cancers, including colon cancer.

How were SSX1, SSX2, and SSX3 expression levels measured?

RT-PCR was used to analyze SSX1, SSX2, and SSX3 expression in 30 matched normal colon (NC) and colon cancer (CC) tissue samples. qRT-PCR was used to assess epigenetic effects in vitro.

What epigenetic treatments were tested and what was the outcome?

CC cells were treated in vitro with 5-aza-20-deoxycytidine (to reduce DNA methyltransferase activity) and trichostatin (to inhibit histone deacetylation). Both treatments significantly induced SSX1, SSX2, and SSX3 mRNA expression in CC cells.

Sep 24, 2026

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