Methylation and mRNA expression levels of P15, death-associated protein kinase, and suppressor of cytokine signaling-1 genes in multiple myeloma

Document Type: Original Article

Authors

1 Department of Hematology, The Second Affiliated Hospital of Kunming Medical University, Kunming 650101, China

2 Department of Hematology, The First Affiliated Hospital of Kunming Medical University, Kunming 650041, China

Abstract

Objective(s): The aim of this study was to investigate the methylation status and mRNA expression levels of P15, death-associated protein kinase (DAPK), and suppressor of cytokine signaling-1 (SOCS1) genes in multiple myeloma (MM).
Materials and Methods: The bone marrow samples of 54 MM patients were collected and the methylation status of the P15, DAPK, and SOCS1 gene promoter regions was determined by methylation-specific polymerase chain reaction. Automated sequencing technology was used to sequence the amplified products in order to analyze the base methylation sites. mRNA expression levels were determined using real-time fluorescent quantitative polymerase chain reaction .
Results: Among the 54 MM patients, the positive methylation rates of the P15, DAPK, and SOCS1 genes were 27.78%, 18.52%, and 16.67%, respectively. The methylation results were confirmed by sequencing. The positive methylation rates of the P15, DAPK, and SOCS1 genes showed no correlation with patient gender, age, typing, staging, and grouping (P>0.05). There was no significant difference in the mRNA expression levels of the P15, DAPK, and SOCS1 genes between the MM patient group and the control group (P>0.05).
Conclusions: Aberrant methylation of the P15, DAPK, and SOCS1 genes exists in MM, and these genes may play certain roles in pathogenesis of MM. There was no significant difference in mRNA expression levels between the methylated group and the non-methylated group, suggesting that these genes are regulated by other mechanisms during their transcription.

Keywords


1. Chapman MA, Lawrence MS, Keats JJ, Cibulskis K, Sougnez C, Schinzel AC, et al. Initial genome sequencing and analysis of multiple myeloma. Nature 2011; 471:467-472.

2. Fonseca R, Bergsagel PL, Drach J, Shaughnessy J, Gutierrez N, Stewart AK, et al. International Myeloma Working Group molecular classification of multiple myeloma: spotlight review. Leukemia 2009; 23:2210-2221.

3. Morgan GJ, Walker BA, Davies FE. The genetic architecture of multiple myeloma. Nat Rev Cancer 2012; 12:335-348.

4. Walker BA, Wardell CP, Chiecchio L, Smith EM, Boyd KD, Neri A, et al. Aerrant global methylation patterns affect the molecular pathogenesis and prognosis of multiple myeloma. Blood 2011; 117:553-562.

5. Raynal NJ, Si J, Taby RF, Gharibyan V, Ahmed S, Jelinek J, et al. DNA methylation does not stably lock gene expression but instead serves as amolecular mark for gene silencing memory. Cancer Res 2012; 72:1170-1181.

6. Jones PA. Functions of DNA methylation: islands, start sites, gene bodies and beyond. Nat Rev Genet 2012; 13:484-492.

7. Dimopoulos K, Gimsing P, Grønbæk K. The role of epigenetics in the biology of multiple myeloma. Blood Cancer J 2014; 4:e207.

8. Li J, Bi L, Lin Y, Lu Z, Hou G. Clinicopathological significance and potential drug target of p15INK4B in multiple myeloma. Drug Des Devel Ther 2014; 8:2129-2136.

9. Kristensen LS, Asmar F, Dimopoulos K, Nygaard MK, Aslan D, Hansen JW, et al. Hypermethylation of DAPK1 is an independent prognostic fator predicting survival in diffuse large B-cell lymphoma. Oncotarget 2014; 5:9798-9810.

10. Collins AS, McCoy CE, Lloyd AT, O'Farrelly C, Stevenson NJ. An effective regulator of SOCS3 and enhancer of JAK-STAT signalling. PLoS One 2013; 8:e69090.

11. Ibanez DE, Caceres I, Cairns P. Methylated DNA sequences for early cancer detection, molecular classification and chemotherapy response prediction. Clin Transl Oncol 2007; 9:429-437.

12. Worthley DL, Whitehall VL, Buttenshaw RL, Irahara N, Greco SA, Ramsnes I, et al. DNA methylation within the normal colorectal mucosa is associated with pathway-specific predisposition to cancer. Oncogene. 2010; 29:1653-1662.

13. Yang B, Guo M, Herman JG, Clark DP. Aberrant promoter methylation profiles of tumor suppressor genes in hepatocellular carcinoma. Am J Pathol 2003; 163:1101-1107.

14. Baylin SB. DNA methylation and gene silencing in cancer. Nat Clin Pract Oncol 2005; 1:S4-11.

15. Amara K, Trimeche M, Ziadi S, Laatiri A, Hachana M, Korbi S. Prognostic significance of aberrant promoter hypermethylation of CpG islands in patients with diffuse large B-cell lymphomas. Ann Oncol 2008; 19:1774-1786.

16. Yang Y, Takeuchi S, Hofmann WK, Ikezoe T, van Dongen JJ, SzczepaƄski T, et al. Aberrant methylation in promoter associated CpG islands of multiple genes in acute lymphoblastic leukemia. Leuk Res 2006; 30:98-102.

17. Chim CS, Fung TK, Cheung WC, Liang R, Kwong YL. SOCS-1 and SHP1 hypermethylation in multiple myeloma: implications for epigenetic activation of the JAK/STAT pathway. Blood. 2004; 103:4630-4635.

18. Galm O, Wilop S, Reichelt J, Jost E, Gehbauer G, Herman JG, et al. DNA methylation changes in multiple myeloma. Leukemia 2004; 18:1687-1692.

19. Dawson MA, Kouzarides T. Cancer epigenetics: from mechanism to therapy. Cell 2012; 150:12-27.

20. Rossi D, Capello D, Gloghini A, Franceschetti S, Paulli M, Bhatia K, et al. Aberrant promoter methylation of multiple genes throughout the clinico-pathologic spectrum of B-cell neoplasia. Haematologica 2004; 89:154-164.

21. Yuregir OO, Yurtcu E, Kizilkilic E, Kocer NE, Ozdogu H, Sahin FI. Detecting methylation patterns of p16, MGMT, DAPK and E-cadherin genes in multiple myeloma patients. Int J Lab Hematol 2010; 32:142-149.

22. Martin P, Garcia-Cosio M, Santon A, Bellas C. Aberrant gene promoter methylation in plasma cell dyscrasias. Exp Mol Pathol 2008; 84:256-261.

23. Esteban B, Angelo M,Maria E. et al. Methylation status  of nine tumor suppressor genes in multiple myeloma. Int J Hematol, 2010,91:87-96

24. Hatzimichael E, Dasoula A, Benetatos L, Syed N, Dranitsaris G, Crook T, et al. Study of specific genetic and epigenetic variables in multiple myeloma. Leuk Lymphoma 2010; 51:2270-2274.

25. Yamamoto M, Nishimoto N, Davydova J, Kishimoto T, Curiel DT. Suppressor of cytokine signaling-1 expression by infectivity-enhanced adenoviral vector inhibits IL-6-dependent proliferation of multiple myeloma cells. Cancer Gene Ther 2006; 13:194-202.

26. Sharma A, Heuck CJ, Fazzari MJ, Mehta J, Singhal S, Greally JM, et al. DNA methylation alterations in multiple myeloma as a model for epigenetic changes in cancer. Wiley Interdiscip Rev Syst Biol Med 2010; 2:654-669.

27. Melzner I, Möller P. Silencing of the SOCS-1 gene by CpG methylation? Blood 2003; 102:1554-1556.

28.  Depil S, Saudemont A, Quesnel B. SOCS-1 gene methylation is frequent but does not appear to have prognostic value in patients with multiple myeloma. Leukemia 2003; 17:1678-1679.

29. Toyooka S, Toyooka KO, Miyajima K, Reddy JL, Toyota M, Sathyanarayana UG, et al. Epigenetic down-regulation of death-associated protein kinase in lung cancers. Clin Cancer Res 2003; 9:3034-3041.

30. Walker BA, Morgan GJ. Could DNA methylation become a useful measure for multiple myeloma prognoses? Expert Rev Hematol 2011; 4:125-127.

31. Shenker N, Flanagan JM. Intragenic DNA methylation: implications of this epigenetic mechanism for cancer research. Br J Cancer 2012; 106:248-253.

32. Jung S, Kim S, Gale M, Cherni I, Fonseca R, Carpten J, et al. DNA Methylation in multiple myeloma is weakly associated with gene transcription. PLoS One 2012; 7:e52626.