Methylation of the MAPK1 Gene as a Potential Diagnostic and Prognostic Biomarker in Childhood B-Acute Lymphoblastic Leukemia
Abstract
Background: Acute lymphoblastic leukemia is a type of heterogeneous leukemia that is associated with the abnormal proliferation of immature monoclonal B and T lymphoid precursors. Cancer is considered an epigenetic disease due to fundamental changes in the pattern of normal methylation in cancer cells. This study aimed to investigate these changes in 5 candidate genes in childhood B cell-ALL.
Materials and Methods: After blood sampling, DNA extraction, and bisulfite treatment, PCR, and Sanger sequencing were performed for part of the TBX3, SLC38A11, SOX2, KCNQ1, and MAPK1 gene promoter. Finally, the methylation pattern of patients in remission and relapse groups was compared with that of healthy individuals.
Results: In the promoter methylation pattern of MAPK1, a significant difference was observed in the CpG3 site between remission-control and relapse-control groups, and in CpG8, 9, 10 between relapse-control and relapse-remission groups. There was no significant difference in the other 4 genes.
Conclusion: We reported the similarity of the methylation pattern in the CpG 3 site of the MAPK1 gene promoter in the remission and relapse groups, as well as the difference in the methylation pattern of this location in the two mentioned groups with control groups, which can suggest the methylation pattern in this site as a candidate for a prognostic biomarker. We also reported the similarity of methylation pattern in remission and control groups on CpG 8,9,10 site and significant reduction of methylation in the relapse group compared to both control and remission groups, which can introduce them as a candidate for diagnostic biomarkers.
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14. Heller G, Topakian T, Altenberger C, et al. Next-generation sequencing identifies major DNA methylation changes during progression of Ph+ chronic myeloid leukemia. Leukemia. 2016;30(9):1861-8.
15. Shen H, Wang Z, Chen Y, et al. Integrative genome-wide aberrant DNA methylation and transcriptome analysis identifies diagnostic markers for colorectal cancer. Arch Toxicol. 2025;99(5):2179-96.
16. Yamazaki J, Jelinek J, Lu Y, et al. TET2 Mutations Affect Non-CpG Island DNA Methylation at Enhancers and Transcription Factor-Binding Sites in Chronic Myelomonocytic Leukemia. Cancer Res. 2015;75(14):2833-43.
17. Li Y, Tollefsbol TO. DNA methylation detection: bisulfite genomic sequencing analysis. Methods Mol Biol. 2011;791:11-21.
18. Reed K, Poulin ML, Yan L, et al. Comparison of bisulfite sequencing PCR with pyrosequencing for measuring differences in DNA methylation. Anal Biochem. 2010;397(1):96-106.
19. Figueroa ME, Chen SC, Andersson AK, P et al. Integrated genetic and epigenetic analysis of childhood acute lymphoblastic leukemia. J Clin Invest. 2013;123(7):3099-111.
20. Pont MJ, Honders MW, et al. Microarray Gene Expression Analysis to Evaluate Cell Type Specific Expression of Targets Relevant for Immunotherapy of Hematological Malignancies. PLoS One. 2016;11(5):e0155165.
21. Arber DA, Orazi A, Hasserjian R, et al. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 2016;127(20):2391-405.
22. Parrish RR, Day JJ, Lubin FD. Direct bisulfite sequencing for examination of DNA methylation with gene and nucleotide resolution from brain tissues. Curr Protoc Neurosci. 2012;Chapter 7:Unit 7.24.
23. Zhang X, Rastogi P, Shah B, et al. B lymphoblastic leukemia/lymphoma: new insights into genetics, molecular aberrations, subclassification and targeted therapy. Oncotarget. 2017;8(39):66728-41.
24. Fang Q, Song Y, Gong X, et al. Gene Deletions and Prognostic Values in B-Linage Acute Lymphoblastic Leukemia. Front Oncol. 2021;11:677034.
25. Huang FL, Liao EC, Li CL, et al. Pathogenesis of pediatric B-cell acute lymphoblastic leukemia: Molecular pathways and disease treatments. Oncol Lett. 2020;20(1):448-54.
26. Rahmani M, Talebi M, Hagh MF, et al. Aberrant DNA methylation of key genes and Acute Lymphoblastic Leukemia. Biomed Pharmacother. 2018;97:1493-500.
27. Sun L, Zhang H, Gao P. Metabolic reprogramming and epigenetic modifications on the path to cancer. Protein Cell. 2022;13(12):877-919.
28. de Barrios O, Parra M. Epigenetic Control of Infant B Cell Precursor Acute Lymphoblastic Leukemia. Int J Mol Sci. 2021;22(6):3127.
29. Ranjbar R, Karimian A, Aghaie Fard A, et al. The importance of miRNAs and epigenetics in acute lymphoblastic leukemia prognosis. J Cell Physiol. 2019;234(4):3216-30.
30. Deng R, Zhang HL, Huang JH, et al. MAPK1/3 kinase-dependent ULK1 degradation attenuates mitophagy and promotes breast cancer bone metastasis. Autophagy. 2021;17(10):3011-29.
31. Li M, Cai O, Yu Y, et al. Paeonol inhibits the malignancy of Apatinib-resistant gastric cancer cells via LINC00665/miR-665/MAPK1 axis. Phytomedicine. 2022;96:153903.
32. Li Y, Luo H, Xiao N, et al. Long Noncoding RNA SChLAP1 Accelerates the Proliferation and Metastasis of Prostate Cancer via Targeting miR-198 and Promoting the MAPK1 Pathway. Oncol Res. 2018;26(1):131-43.
33. Lu M, Gao Q, Wang Y, et al. LINC00511 promotes cervical cancer progression by regulating the miR-497-5p/MAPK1 axis. Apoptosis. 2022;27(11-12):800-11.
34. Najar AG, Pashaei-Asl R, Omidi Y, et al. EGFR antisense oligonucleotides encapsulated with nanoparticles decrease EGFR, MAPK1 and STAT5 expression in a human colon cancer cell line. Asian Pac J Cancer Prev. 2013;14(1):495-8.
35. Scheffel RS, Dora JM, Maia AL. BRAF mutations in thyroid cancer. Curr Opin Oncol. 2022;34(1):9-18.
36. Xu M, Li J, Wang X, et al. MiR-22 suppresses epithelial-mesenchymal transition in bladder cancer by inhibiting Snail and MAPK1/Slug/vimentin feedback loop. Cell Death Dis. 2018;9(2):209.
37. Yan Z, Ohuchida K, Fei S, et al. Inhibition of ERK1/2 in cancer-associated pancreatic stellate cells suppresses cancer-stromal interaction and metastasis. J Exp Clin Cancer Res. 2019;38(1):221.
38. Kageyama R, Pastan I. Molecular cloning and characterization of a human DNA binding factor that represses transcription. Cell. 1989;59(5):815-25.
39. Yang C, Yin L, Zhou P, et al. Transcriptional regulation of IER5 in response to radiation in HepG2. Cancer Gene Ther. 2016;23(2-3):61-5.
40. Gondim MV, Wiltzer-Bach L, Maurer B, et al. AP-2 Is the Crucial Clathrin Adaptor Protein for CD4 Downmodulation by HIV-1 Nef in Infected Primary CD4+ T Cells. J Virol. 2015;89(24):12518-24.
2. Sindik N, Pereza N, Dević Pavlić S. Epigenetics of oogenesis. Arch Gynecol Obstet. 2025;311(2):183-90.
3. Ferrer J, Dimitrova N. Transcription regulation by long non-coding RNAs: mechanisms and disease relevance. Nat Rev Mol Cell Biol. 2024;25(5):396-415.
4. Tejedor JR, Bueno C, Vinyoles M, et al. Integrative methylome-transcriptome analysis unravels cancer cell vulnerabilities in infant MLL-rearranged B cell acute lymphoblastic leukemia. J Clin Invest. 2021;131(13):e138833.
5. Seong H, Izutsu R, Osaki M, et al. Cancer prevention: past challenges and future directions. Genes Environ. 2025;47(1):4.
6. Tran TH, Tasian SK. How I treat Philadelphia chromosome-like acute lymphoblastic leukemia in children, adolescents, and young adults. Blood. 2025;145(1):20-34.
7. Du Y, Zhang X, Sun M, et al. Molecular characterization and biomarker identification in paediatric B-cell acute lymphoblastic leukaemia. J cell Mol Med. 2024;28(19):e70126.
8. Lei S, Jia S, Takalkar S, et al. Genomic profiling of circulating tumor DNA for childhood cancers. Leukemia. 2025;39(2):420-30.
9. Mullighan CG. Molecular genetics of B-precursor acute lymphoblastic leukemia. J Clin Invest. 2012;122(10):3407-15.
10. Xu H, Yu H, Jin R, et al. Genetic and Epigenetic Targeting Therapy for Pediatric Acute Lymphoblastic Leukemia. Cells. 2021;10(12):3349.
11. Nordlund J, Syvänen AC. Epigenetics in pediatric acute lymphoblastic leukemia. Semin Cancer Biol. 2018;51:129-38.
12. Lee Y, Dho SH, Lee J, et al. Hypermethylation of PDX1, EN2, and MSX1 predicts the prognosis of colorectal cancer. Exp Mol Med. 2022;54(2):156-68.
13. Neja SA. Site-Specific DNA Demethylation as a Potential Target for Cancer Epigenetic Therapy. Epigenet Insights. 2020;13:2516865720964808.
14. Heller G, Topakian T, Altenberger C, et al. Next-generation sequencing identifies major DNA methylation changes during progression of Ph+ chronic myeloid leukemia. Leukemia. 2016;30(9):1861-8.
15. Shen H, Wang Z, Chen Y, et al. Integrative genome-wide aberrant DNA methylation and transcriptome analysis identifies diagnostic markers for colorectal cancer. Arch Toxicol. 2025;99(5):2179-96.
16. Yamazaki J, Jelinek J, Lu Y, et al. TET2 Mutations Affect Non-CpG Island DNA Methylation at Enhancers and Transcription Factor-Binding Sites in Chronic Myelomonocytic Leukemia. Cancer Res. 2015;75(14):2833-43.
17. Li Y, Tollefsbol TO. DNA methylation detection: bisulfite genomic sequencing analysis. Methods Mol Biol. 2011;791:11-21.
18. Reed K, Poulin ML, Yan L, et al. Comparison of bisulfite sequencing PCR with pyrosequencing for measuring differences in DNA methylation. Anal Biochem. 2010;397(1):96-106.
19. Figueroa ME, Chen SC, Andersson AK, P et al. Integrated genetic and epigenetic analysis of childhood acute lymphoblastic leukemia. J Clin Invest. 2013;123(7):3099-111.
20. Pont MJ, Honders MW, et al. Microarray Gene Expression Analysis to Evaluate Cell Type Specific Expression of Targets Relevant for Immunotherapy of Hematological Malignancies. PLoS One. 2016;11(5):e0155165.
21. Arber DA, Orazi A, Hasserjian R, et al. The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood. 2016;127(20):2391-405.
22. Parrish RR, Day JJ, Lubin FD. Direct bisulfite sequencing for examination of DNA methylation with gene and nucleotide resolution from brain tissues. Curr Protoc Neurosci. 2012;Chapter 7:Unit 7.24.
23. Zhang X, Rastogi P, Shah B, et al. B lymphoblastic leukemia/lymphoma: new insights into genetics, molecular aberrations, subclassification and targeted therapy. Oncotarget. 2017;8(39):66728-41.
24. Fang Q, Song Y, Gong X, et al. Gene Deletions and Prognostic Values in B-Linage Acute Lymphoblastic Leukemia. Front Oncol. 2021;11:677034.
25. Huang FL, Liao EC, Li CL, et al. Pathogenesis of pediatric B-cell acute lymphoblastic leukemia: Molecular pathways and disease treatments. Oncol Lett. 2020;20(1):448-54.
26. Rahmani M, Talebi M, Hagh MF, et al. Aberrant DNA methylation of key genes and Acute Lymphoblastic Leukemia. Biomed Pharmacother. 2018;97:1493-500.
27. Sun L, Zhang H, Gao P. Metabolic reprogramming and epigenetic modifications on the path to cancer. Protein Cell. 2022;13(12):877-919.
28. de Barrios O, Parra M. Epigenetic Control of Infant B Cell Precursor Acute Lymphoblastic Leukemia. Int J Mol Sci. 2021;22(6):3127.
29. Ranjbar R, Karimian A, Aghaie Fard A, et al. The importance of miRNAs and epigenetics in acute lymphoblastic leukemia prognosis. J Cell Physiol. 2019;234(4):3216-30.
30. Deng R, Zhang HL, Huang JH, et al. MAPK1/3 kinase-dependent ULK1 degradation attenuates mitophagy and promotes breast cancer bone metastasis. Autophagy. 2021;17(10):3011-29.
31. Li M, Cai O, Yu Y, et al. Paeonol inhibits the malignancy of Apatinib-resistant gastric cancer cells via LINC00665/miR-665/MAPK1 axis. Phytomedicine. 2022;96:153903.
32. Li Y, Luo H, Xiao N, et al. Long Noncoding RNA SChLAP1 Accelerates the Proliferation and Metastasis of Prostate Cancer via Targeting miR-198 and Promoting the MAPK1 Pathway. Oncol Res. 2018;26(1):131-43.
33. Lu M, Gao Q, Wang Y, et al. LINC00511 promotes cervical cancer progression by regulating the miR-497-5p/MAPK1 axis. Apoptosis. 2022;27(11-12):800-11.
34. Najar AG, Pashaei-Asl R, Omidi Y, et al. EGFR antisense oligonucleotides encapsulated with nanoparticles decrease EGFR, MAPK1 and STAT5 expression in a human colon cancer cell line. Asian Pac J Cancer Prev. 2013;14(1):495-8.
35. Scheffel RS, Dora JM, Maia AL. BRAF mutations in thyroid cancer. Curr Opin Oncol. 2022;34(1):9-18.
36. Xu M, Li J, Wang X, et al. MiR-22 suppresses epithelial-mesenchymal transition in bladder cancer by inhibiting Snail and MAPK1/Slug/vimentin feedback loop. Cell Death Dis. 2018;9(2):209.
37. Yan Z, Ohuchida K, Fei S, et al. Inhibition of ERK1/2 in cancer-associated pancreatic stellate cells suppresses cancer-stromal interaction and metastasis. J Exp Clin Cancer Res. 2019;38(1):221.
38. Kageyama R, Pastan I. Molecular cloning and characterization of a human DNA binding factor that represses transcription. Cell. 1989;59(5):815-25.
39. Yang C, Yin L, Zhou P, et al. Transcriptional regulation of IER5 in response to radiation in HepG2. Cancer Gene Ther. 2016;23(2-3):61-5.
40. Gondim MV, Wiltzer-Bach L, Maurer B, et al. AP-2 Is the Crucial Clathrin Adaptor Protein for CD4 Downmodulation by HIV-1 Nef in Infected Primary CD4+ T Cells. J Virol. 2015;89(24):12518-24.
| Files | ||
| Issue | Vol 20 No 3 (2026) | |
| Section | Original Article(s) | |
| Keywords | ||
| Acute lymphoblastic leukemia; Bisulfite sanger sequencing; Epigenetics; MAPK1; Promoter-methylation | ||
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How to Cite
1.
Rajabian Z, Koohini Z. Methylation of the MAPK1 Gene as a Potential Diagnostic and Prognostic Biomarker in Childhood B-Acute Lymphoblastic Leukemia. Int J Hematol Oncol Stem Cell Res. 2026;20(3):257-267.
