Journal of Rafsanjan University of Medical Sciences
Volume 25, Issue 4 (7-2026)
JRUMS 2026, 25(4): 413-432 |
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Ethics code: IR.RUMS.REC.1399.261
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Eshkevari S M S, Bagheri-Hosseinabadi Z, Khalighfard S, Mahmoodi M, Alizadeh A M. Evaluation of DNA Methylation Changes in CTNNB1 and GPRASP1 Genes in the Signaling Pathway of Interleukins 4 and 13 as Radiotherapy Resistance Biomarkers in Serum Cell-Free DNA of Patients with Non-Metastatic Rectal Cancer: A Bioinformatics and Experimental Study. JRUMS 2026; 25 (4) :413-432
URL: http://journal.rums.ac.ir/article-1-7535-en.html
URL: http://journal.rums.ac.ir/article-1-7535-en.html
Seyed Mohammad Sadat Eshkevari 
, Zahra Bagheri-Hosseinabadi , Solmaz Khalighfard , Mehdi Mahmoodi , Ali Mohammad Alizadeh *
, Zahra Bagheri-Hosseinabadi , Solmaz Khalighfard , Mehdi Mahmoodi , Ali Mohammad Alizadeh *
Cancer Research Center, Cancer Institute, Tehran University of Medical Sciences, Tehran, Iran
Abstract: (15 Views)
Background and Objectives: Inflammation and epigenetic changes are considered major factors in developing rectal cancer. The aim of this study was to determine methylation changes of CTNNB1 and GPRASP1 genes in the signaling pathway of interleukins 4 and 13 in patients with non-metastatic rectal cancer.
Materials and Methods: In this quasi-experimental study, bioinformatics analysis was first performed using GO, UALCAN, and DNMIVD databases to identify signaling pathways of target genes. Subsequently, blood samples were collected from 43 patients with rectal cancer (before and after radiotherapy) and 10 healthy individuals. We determined methylation changes in the CTNNB1 and GPRASP1 genes by methylation-specific PCR and measured serum levels of interleukins 4 and 13 by ELISA. We analyzed parametric data using an independent t-test and non-parametric data using the Mann-Whitney U test.
Results: In bioinformatics analysis, CTNNB1 was identified as a hypomethylated gene (with high expression), and GPRASP1 was identified as a hypermethylated gene (with low expression). After radiotherapy, increased methylation of CTNNB1 led to a significant decrease in its expression, while decreased methylation of GPRASP1 resulted in a significant increase in its expression (P<0.05). Moreover, serum levels of interleukin-13 significantly decreased, and that of interleukin-4 significantly increased after radiotherapy compared to before (P<0.05).
Conclusion: The results of the current study demonstrated that radiotherapy, by inducing changes in DNA methylation accompanied by alterations in serum levels of interleukins 4 and 13, could serve as an effective model for monitoring treatment response in patients with non-metastatic rectal cancer.
Keywords: Rectal cancer, Cell-free DNA, DNA methylation, Radiotherapy, Interleukin 4, Interleukin 13
Funding: This study was co-funded by Tehran University of Medical Sciences and Rafsanjan University of Medical Sciences.
Conflict of interest: None declared.
Ethical approval: The Ethics Committee of Tehran University of Medical Sciences (IR.TUMS.IKHC.REC.1399.320) and Rafsanjan University of Medical Sciences (IR.RUMS.REC.1399.261) approved the study.
Authors’ contributions
Conceptualization: Ali Mohammad Alizadeh, Zahra Bagheri-Hosseinabadi, Solmaz Khalighfard
Methodology: Seyed Mohammad Sadat Eshkevari, Solmaz Khalighfard
Data collection: Seyed Mohammad Sadat Eshkevari, Solmaz Khalighfard
Formal analysis: Solmaz Khalighfard
Supervision: Zahra Bagheri-Hosseinabadi, Mehdi Mahmoodi
Project administration: Ali Mohammad Alizadeh
Writing – original draft: Seyed Mohammad Sadat Eshkevari, Solmaz Khalighfard
Writing – review & editing: Zahra Bagheri-Hosseinabadi, Ali Mohammad Alizadeh
Materials and Methods: In this quasi-experimental study, bioinformatics analysis was first performed using GO, UALCAN, and DNMIVD databases to identify signaling pathways of target genes. Subsequently, blood samples were collected from 43 patients with rectal cancer (before and after radiotherapy) and 10 healthy individuals. We determined methylation changes in the CTNNB1 and GPRASP1 genes by methylation-specific PCR and measured serum levels of interleukins 4 and 13 by ELISA. We analyzed parametric data using an independent t-test and non-parametric data using the Mann-Whitney U test.
Results: In bioinformatics analysis, CTNNB1 was identified as a hypomethylated gene (with high expression), and GPRASP1 was identified as a hypermethylated gene (with low expression). After radiotherapy, increased methylation of CTNNB1 led to a significant decrease in its expression, while decreased methylation of GPRASP1 resulted in a significant increase in its expression (P<0.05). Moreover, serum levels of interleukin-13 significantly decreased, and that of interleukin-4 significantly increased after radiotherapy compared to before (P<0.05).
Conclusion: The results of the current study demonstrated that radiotherapy, by inducing changes in DNA methylation accompanied by alterations in serum levels of interleukins 4 and 13, could serve as an effective model for monitoring treatment response in patients with non-metastatic rectal cancer.
Keywords: Rectal cancer, Cell-free DNA, DNA methylation, Radiotherapy, Interleukin 4, Interleukin 13
Funding: This study was co-funded by Tehran University of Medical Sciences and Rafsanjan University of Medical Sciences.
Conflict of interest: None declared.
Ethical approval: The Ethics Committee of Tehran University of Medical Sciences (IR.TUMS.IKHC.REC.1399.320) and Rafsanjan University of Medical Sciences (IR.RUMS.REC.1399.261) approved the study.
Authors’ contributions
Conceptualization: Ali Mohammad Alizadeh, Zahra Bagheri-Hosseinabadi, Solmaz Khalighfard
Methodology: Seyed Mohammad Sadat Eshkevari, Solmaz Khalighfard
Data collection: Seyed Mohammad Sadat Eshkevari, Solmaz Khalighfard
Formal analysis: Solmaz Khalighfard
Supervision: Zahra Bagheri-Hosseinabadi, Mehdi Mahmoodi
Project administration: Ali Mohammad Alizadeh
Writing – original draft: Seyed Mohammad Sadat Eshkevari, Solmaz Khalighfard
Writing – review & editing: Zahra Bagheri-Hosseinabadi, Ali Mohammad Alizadeh
Keywords: Rectal cancer, Cell-free DNA, DNA methylation, Radiotherapy, Interleukin 4, Interleukin 13
Type of Study: Research |
Subject:
ايمونولوژي
Received: 2024/10/16 | Accepted: 2026/09/26 | Published: 2026/07/31
Received: 2024/10/16 | Accepted: 2026/09/26 | Published: 2026/07/31
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