Aberrant DNA Methylation Status and mRNA Expression Level of SMG1 Gene in Chronic Myeloid Leukemia: A Case-Control Study

Document Type : Original Article

Authors

1 Department of Hematology and Blood Transfusion, Students Research Center, School of Allied Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Immunology, Pasteur Institute of Iran, Tehran, Iran

3 Department of Medical Biotechnology, School of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, Iran

4 Hematologic Malignancies Research Center, Tehran University of Medical Sciences, Tehran, Iran

5 Department of Medical Laboratory Sciences, School of Paramedicine, Qazvin University of Medical Sciences, Qazvin, Iran

Abstract

Objective
Chronic myeloid leukemia (CML) is a myeloproliferative malignancy with different stages. Aberrant epigenetic
modifications, such as DNA methylation, have been introduced as a signature for diverse cancers which also plays a
crucial role in CML pathogenesis and development. Suppressor with morphogenetic effect on genitalia (SMG1) gene
recently has been brought to the spotlight as a potent tumor suppressor gene that can be suppressed by tumors for
further progress. The present study aims to investigate SMG1 status in CML patients.

Materials and Methods
In this case-control study, peripheral blood from 30 patients with different phases of CML [new
case (N)=10, complete molecular remission (CMR)=10, blastic phase (BP)=10] and 10 healthy subjects were collected.
Methylation status and expression level of SMG1 gene promoter was assessed by methylation-specific polymerase
chain reaction (MSP) and quantitative reverse-transcription PCR, respectively.

Results
MSP results of SMG1 gene promotor in the new case group were methylated (60% methylated, 30%
hemimethylated and 10% unmethylated). All CMR and control group patients were unmethylated in the SMG1 gene
promoter. In the BP group, methylated SMG1 promoter was seen (50% of patients had a methylated status and 50%
had hemimethylated status). In comparison with the healthy subjects, expression level of SMG1 in the new case group
was decreased (P<0.01); in the CMR group and BP-CML groups, it was increased (P<0.05). No significant correlation
between patients’ hematological features and SMG1 methylation was seen.

Conclusion
Our results demonstrated that aberrant methylation of SMG1 occurred in CML patients and it had a
significant association with SMG1 expression. SMG1 gene promoter showed diverse methylated status and subsequent
expression levels in different phases of CML. These findings suggested possible participation of SMG1 suppression in
the CML pathogenesis.

Keywords


1.Roman-Gomez J, Jimenez-Velasco A, Agirre X, Castillejo JA, Navarro G, Jose-Eneriz ES, et al. Epigenetic regulation of PRAME gene in chronic myeloid leukemia. Leuk Res. 2007; 31(11): 1521-1528.
2. Kamachi K, Ureshino H, Watanabe T, Yoshida N, Yamamoto Y, Kurahashi Y, et al. Targeting DNMT1 by demethylating agent OR-2100 increases tyrosine kinase inhibitors-sensitivity and depletes leukemic stem cells in chronic myeloid leukemia. Cancer Lett. 2022; 526: 273-283.
3. How J, Venkataraman V, Hobbs GS. Blast and accelerated phase CML: room for improvement. Hematology Am Soc Hematol Educ Program. 2021; 2021(1): 122-128.
4. Calabretta B, Perrotti D. The biology of CML blast crisis. Blood. 2004; 103(11): 4010-4022.
5. Celik S, Akcora D, Ozkan T, Varol N, Aydos S, Sunguroglu A. Methylation analysis of the DAPK1 gene in imatinib-resistant chronic myeloid leukemia patients. Oncol Lett. 2015; 9(1): 399-404.
6. Wang YL, Qian J, Lin J, Yao DM, Qian Z, Zhu ZH, et al. Methylation status of DDIT3 gene in chronic myeloid leukemia. J Exp Clin Cancer Res. 2010; 29(1): 54.
7. Yao DM, Zhou JD, Zhang YY, Yang L, Wen XM, Yang J, et al. GPX3 promoter is methylated in chronic myeloid leukemia. Int J Clin Exp Pathol. 2015; 8(6): 6450-6457.
8. Jelinek J, Gharibyan V, Estecio MR, Kondo K, He R, Chung W, et al. Aberrant DNA methylation is associated with disease progression, resistance to imatinib and shortened survival in chronic myelogenous leukemia. PLoS One. 2011; 6(7): e22110.
9. Dunwell T, Hesson L, Rauch TA, Wang L, Clark RE, Dallol A, et al. A genome-wide screen identifies frequently methylated genes in haematological and epithelial cancers. Mol Cancer. 2010; 9: 44.
10. Amabile G, Di Ruscio A, Müller F, Welner RS, Yang H, Ebralidze AK, et al. Dissecting the role of aberrant DNA methylation in human leukaemia. Nat Commun. 2015; 6: 7091.
11. Annamaneni S, Kagita S, Gorre M, Digumarti RR, Satti V, Battini MR. Methylation status of CEBPA gene promoter in chronic myeloid leukemia. Hematology. 2014; 19(1): 42-44.
12. Al-Jamal HA, Jusoh SA, Yong AC, Asan JM, Hassan R, Johan MF. Silencing of suppressor of cytokine signaling-3 due to methylation results in phosphorylation of STAT3 in imatinib resistant BCR-ABL positive chronic myeloid leukemia cells. Asian Pac J Cancer Prev.2014; 15(11): 4555-4561.
13. Alipour S, Sakhinia E, Khabbazi A, Samadi N, Babaloo Z, Azad M, et al. Methylation status of interleukin-6 gene promoter in patients with Behçet’s disease. Reumatol Clin (Engl Ed). 2020; 16(3): 229-234.
14. Roman-Gomez J, Jimenez-Velasco A, Agirre X, Castillejo JA, Navarro G, San Jose-Eneriz E, et al. Repetitive DNA hypomethylation in the advanced phase of chronic myeloid leukemia. Leuk Res. 2008; 32(3): 487-490.
15. Yamashita A, Ohnishi T, Kashima I, Taya Y, Ohno S. Human SMG-1, a novel phosphatidylinositol 3-kinase-related protein kinase, associates with components of the mRNA surveillance complex and is involved in the regulation of nonsense-mediated mRNA decay. Genes Dev. 2001; 15(17): 2215-2228.
16. Gubanova E, Brown B, Ivanov SV, Helleday T, Mills GB, Yarbrough WG, et al. Downregulation of SMG-1 in HPV-positive head and neck squamous cell carcinoma due to promoter hypermethylation correlates with improved survival. Clin Cancer Res. 2012; 18(5): 1257-1267.
17. Du Y, Lu F, Li P, Ye J, Ji M, Ma D, et al. SMG1 acts as a novel potential tumor suppressor with epigenetic inactivation in acute myeloid leukemia. Int J Mol Sci. 2014; 15(9): 17065-17076.
18. Rebeiro P, James A, Ling S, Caxeiro N, Lee CS, Roberts T. The role of SMG1 expression in B cell lymphoproliferative disease. Pathology. 2016; 48(1): S74.
19. Figueroa ME, Lugthart S, Li Y, Erpelinck-Verschueren C, Deng X, Christos PJ, et al. DNA methylation signatures identify biologically distinct subtypes in acute myeloid leukemia. Cancer Cell. 2010; 17(1): 13-27.
20. Akalin A, Garrett-Bakelman FE, Kormaksson M, Busuttil J, Zhang L, Khrebtukova I, et al. Base-pair resolution DNA methylation sequencing reveals profoundly divergent epigenetic landscapes in acute myeloid leukemia. PLoS Genet. 2012; 8(6): e1002781.
21. Aslani S, Mahmoudi M, Garshasbi M, Jamshidi AR, Karami J, Nicknam MH. Evaluation of DNMT1 gene expression profile and methylation of its promoter region in patients with ankylosing spondylitis. Clin Rheumatol. 2016; 35(11): 2723-2731.
22. Kanwal R, Gupta S. Epigenetic modifications in cancer. Clin Genet. 2012; 81(4): 303-311.
23. Mencalha AL, Corrêa S, Salles D, Du Rocher B, Santiago MF, Abdelhay E. Inhibition of STAT3-interacting protein 1 (STATIP1) promotes STAT3 transcriptional up-regulation and imatinib mesylate resistance in the chronic myeloid leukemia. BMC Cancer. 2014; 14: 866.
24. Lloyd JP, Davies B. SMG1 is an ancient nonsense-mediated mRNA decay effector. Plant J. 2013; 76(5): 800-810.
25. McIlwain DR, Pan Q, Reilly PT, Elia AJ, McCracken S, Wakeham AC, et al. Smg1 is required for embryogenesis and regulates diverse genes via alternative splicing coupled to nonsense-mediated mRNA decay. Proc Natl Acad Sci USA. 2010; 107(27): 12186-12191.
26. Horejsí Z, Takai H, Adelman CA, Collis SJ, Flynn H, Maslen S, etal. CK2 phospho-dependent binding of R2TP complex to TEL2 is essential for mTOR and SMG1 stability. Mol Cell. 2010; 39(6): 839-850.
27. González-Estévez C, Felix DA, Smith MD, Paps J, Morley SJ, James V, et al. SMG-1 and mTORC1 act antagonistically to regulate response to injury and growth in planarians. PLoS Genet. 2012; 8(3): e1002619.
28. Chen RQ, Yang QK, Chen YL, Oliveira VA, Dalton WS, Fearns C, et al. Kinome siRNA screen identifies SMG-1 as a negative regulator of hypoxia-inducible factor-1alpha in hypoxia. J Biol Chem. 2009; 284(25): 16752-16758.
29. Cheung HH, St Jean M, Beug ST, Lejmi-Mrad R, LaCasse E, Baird SD, et al. SMG1 and NIK regulate apoptosis induced by Smac mimetic compounds. Cell Death Dis. 2011; 2(4): e146.
30. Gubanova E, Issaeva N, Gokturk C, Djureinovic T, Helleday T. SMG-1 suppresses CDK2 and tumor growth by regulating both the p53 and Cdc25A signaling pathways. Cell Cycle. 2013; 12(24): 3770-3780.
31. Wong C, Chen F, Alirezaie N, Wang Y, Cuggia A, Borgida A, et al. A region-based gene association study combined with a leaveone- out sensitivity analysis identifies SMG1 as a pancreatic cancer susceptibility gene. PLoS Genet. 2019; 15(8): e1008344.
32. Han LL, Nan HC, Tian T, Guo H, Hu TH, Wang WJ, et al. Expression and significance of the novel tumor-suppressor gene SMG-1 in hepatocellular carcinoma. Oncol Rep. 2014; 31(6): 2569-2578.
33. Mai S, Xiao R, Shi L, Zhou X, Yang T, Zhang M, et al. MicroRNA-18a promotes cancer progression through SMG1 suppression and mTOR pathway activation in nasopharyngeal carcinoma. Cell Death Dis. 2019; 10(11): 819.
34. Zeng S, Liu S, Feng J, Gao J, Xue F. MicroRNA-32 promotes ovarian cancer cell proliferation and motility by targeting SMG1. Oncol Lett. 2020; 20(1): 733-741.
35. Zhang X, Peng Y, Huang Y, Yang M, Yan R, Zhao Y, et al. SMG-1 inhibition by miR-192/-215 causes epithelial-mesenchymal transition in gastric carcinogenesis via activation of Wnt signaling. Cancer Med. 2018; 7(1): 146-156.
36. Ding X, Yang Y, Sun Y, Xu W, Su B, Zhou X. MicroRNA-585 acts as a tumor suppressor in non-small-cell lung cancer by targeting hSMG-1. Clin Transl Oncol. 2017; 19(5): 546-552.
37. Yin Z, Ma T, Yan J, Shi N, Zhang C, Lu X, et al. LncRNA MAGI2-AS3 inhibits hepatocellular carcinoma cell proliferation and migration by targeting the miR-374b-5p/SMG1 signaling pathway. J Cell Physiol. 2019; 234(10): 18825-18836.
38. Zhang Y, Zheng Y, Faheem A, Sun T, Li C, Li Z, et al. A novel AKT inhibitor, AZD5363, inhibits phosphorylation of AKT downstream molecules, and activates phosphorylation of mTOR and SMG-1 dependent on the liver cancer cell type. Oncol Lett. 2016; 11(3): 1685-1692.
39. Sun S, Fang H. Curcumin inhibits ovarian cancer progression by regulating circ-PLEKHM3/miR-320a/SMG1 axis. J Ovarian Res. 2021; 14(1): 158.