Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Suppression of DNMTs Accelerates the In Vitro Erythropoietic Differentiation of Human $CD34^+$ Progenitor Cells

  • Kim, Seok-Ho (Department of Oral Biochemistry, The 2nd Stage of BK21 Program for Dental School, Dental Science Research Institute, Chonnam National University) ;
  • Yang, Hee-Young (Department of Oral Biochemistry, The 2nd Stage of BK21 Program for Dental School, Dental Science Research Institute, Chonnam National University) ;
  • Jeong, Dong-Kee (Department of Animal Science & Biotechnology, College of Applied Life Science, Cheju National University) ;
  • Lee, Sang-Ryeul (Department of Oral Biochemistry, The 2nd Stage of BK21 Program for Dental School, Dental Science Research Institute, Chonnam National University) ;
  • Ryoo, Zae-Young (School of Life Science & Biotechnology, Kyungpook National University) ;
  • Lee, Tae-Hoon (Department of Oral Biochemistry, The 2nd Stage of BK21 Program for Dental School, Dental Science Research Institute, Chonnam National University)
  • Published : 2007.12.31
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Abstract

Epigenetic modification dependent DNA methyltransferases (DNMTs) play an important role in tissue- and stage-specific gene regulation and normal mammalian development. In this study, we show that DNMTs are expressed at different levels during hematopoietic stem cell (HSC) differentiation to proerythrocytes. DNMT1, DNMT3A, and DNMT3B were highly expressed at day 7 after differentiation. We used specific siRNA as a tool to probe the relationship between the expression of DNMTs and erythropoietic differentiation. When introduced siRNA of DMNT1 and DMNT3b in human $CD34^+$ cells, these more differentiated into erythrocytes. This was confirmed by glycophorin A (GPA) positive cell analysis and globin gene expression. $GPA^+$ cells increased up to $20{\sim}30%$, and ${\gamma}$- and ${\epsilon}$-globin genes increased in siRNA transfected cells. Therefore, our data suggest that suppression of DNA methylation can affect positively differentiation of HSC and may contribute to expression of erythrocyte lineage genes including GPA and globins.

Keywords

References

  1. Baiocchi M, Di Rico C, Di Pietro R, Di Baldassarre A, Migliaccio AR (2003): 5-azacytidine reactivates the erythroid differentiation potential of the myeloid-restricted murine cell line 32D Ro. Exp Cell Res 285:258-267 https://doi.org/10.1016/S0014-4827(03)00032-6
  2. Bestor TH (2000): The DNA methyltransferases of mammals. Hum Mol Genet 9:2395-2402 https://doi.org/10.1093/hmg/9.16.2395
  3. Bird A (2002): DNA methylation patterns and epigenetic memory. Gene Dev 16:6-21 https://doi.org/10.1101/gad.947102
  4. Cao H, Stamatoyannopoulos G, Jung M (2004): Induction of human gamma globin gene expression by histone deacetylase inhibitors. Blood 103:701-709 https://doi.org/10.1182/blood-2003-02-0478
  5. Felgner J, Heidorn K, Korbacher D, Frahm SO, Parwaresch R (1999): Cell lineage specificity in G-CSF receptor gene methylation. Leukemia 13:530-534 https://doi.org/10.1038/sj/leu/2401386
  6. Gomes I, Sharma TI, Edassery S, Fulton N, Mar BG, Westbrook CA (2002): Novel transcription factors in human CD34 antigen-positive hematopoietic cells. Blood 100:107-119 https://doi.org/10.1182/blood.V100.1.107
  7. Laird PW (2005): Cancer epigenetics. Hum Mol Genet 14(Sl):R65-76 https://doi.org/10.1093/hmg/ddi113
  8. Leonhardt H, Page AW, Weier HO, Bestor TH (1992): A targeting sequence directs DNA methyltransferase to sites of DNA replication in mammalian nuclei. Cell 71: 865-873 https://doi.org/10.1016/0092-8674(92)90561-P
  9. Leu YW, Rahmatpanah F, Shi H, Wei SH, Liu JC, Yan PS, Huang TH (2003): Double RNA interference of DNMT3b and DNMT1 enhances DNA demethylation and gene reactivation. Cancer Res 63: 6110-6115
  10. Li LH, Olin EJ, Buskirk HH, Reineke LM (1970): Cytotoxicity and mode of action of 5-azaeytidine on L1210 leukemia. Cancer Res 30:2760-2769
  11. Lubbert M, Miller CW, Koeffler HP (1991): Changes of DNA methylation and chromatin structure in the human myeloperoxidase gene during myeloid differentiation. Blood 78:345-356
  12. Maraschio P, Zuffardi O, Dalla Fior T, Tiepolo L (1988): Immunodeficiency, centromeric heterochromatin instability of chromosomes 1, 9, and 16, and facial anomalies: the ICF syndrome. J Med Genet 25:173-180 https://doi.org/10.1136/jmg.25.3.173
  13. Milhem M, Mahmud N, Lavelle D, Araki I-L DeSimone J, Saunthararajah Y, Hoffman R (2004): Modification of hematopoietic stem cell fate by 5-aza 2'deoxyeytidine and trichostatin A. Blood 103:4102-4110 https://doi.org/10.1182/blood-2003-07-2431
  14. Mizuno S, Chijiwa T, Okamura T, Akashi K, Fukumaki Y, Niho Y, Sasaki H (2001): Expression of DNA methyltransferases DNMTl, 3A, and 3B in normal hematopoiesis and in acute and chronic myelogenous leukemia. Blood 97(5):1172-1179 https://doi.org/10.1182/blood.V97.5.1172
  15. Okano M, Bell DW, Haber DA, Li DA (1999): DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 99:247-257 https://doi.org/10.1016/S0092-8674(00)81656-6
  16. Oridate N, Lotan R (2005): Suppression of DNA methyltransferase 1 levels in head and neck squamous carcinoma cells using small interfering RNA results in growth inhibition and increase in Cdk inhibitor p21. Int J Onco 26:757-761
  17. Saunthararajah Y, Hillery CA, Lavelle D, Molokie R, Dom L (2003): Effects of 5-aza-2'-deoxyeytidine on fetal hemoglobin levels, red cell adhesion, and hematopoietic differentiation in patients with sickle cell disease. Blood 102:3865-3870 https://doi.org/10.1182/blood-2003-05-1738
  18. Saunthararajah Y, Lavelle D, DeSimone J (2004): DNA hypo-methylating agents and sickle cell disease. Brit J Haematol 126:629-636 https://doi.org/10.1111/j.1365-2141.2004.05064.x
  19. Stamatoyannopoulos G (2005): Control of globin gene expression during development and erythroid differentiation. Exp Hematol 33:259-271 https://doi.org/10.1016/j.exphem.2004.11.007
  20. Suzuki M,. Sunaga N, Shames DS, Toyooka S, Gazdar AF, Minna JD (2004): RNA interference-mediated knockdown of DNA methyltransferase 1 leads to promoter demethylation and gene re-expression in human lung and breast cancer cells. Cancer Res 64:3137-3143 https://doi.org/10.1158/0008-5472.CAN-03-3046
  21. Yang HY, Kim SH, Kim SH, Kim DJ, Kim SU, Yu DY, Yeom Yl, Lee DS, Kim YJ, Park BJ, Lee TH (2007): The suppression of zfpm-1 accelerates the erythropoietic differentiation of human $CD34^+$ cells. Biochem Biophys Res Commun 353(4):978-984 https://doi.org/10.1016/j.bbrc.2006.12.155
  22. Yin H, Blanchard KL (2000): DNA methylation represses the expression of the human erythropoietin gene by two different mechanisms. Blood 95:111-119