Browse > Article

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)
Publication Information
Reproductive and Developmental Biology / v.31, no.4, 2007 , pp. 241-248 More about this Journal
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
Erythropoiesis; DNA methyltransferase; Hematopoietic stem cell; In vitro differentiation; siRNA;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Felgner J, Heidorn K, Korbacher D, Frahm SO, Parwaresch R (1999): Cell lineage specificity in G-CSF receptor gene methylation. Leukemia 13:530-534   DOI   ScienceOn
2 Laird PW (2005): Cancer epigenetics. Hum Mol Genet 14(Sl):R65-76   DOI   ScienceOn
3 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
4 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   DOI
5 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
6 Saunthararajah Y, Lavelle D, DeSimone J (2004): DNA hypo-methylating agents and sickle cell disease. Brit J Haematol 126:629-636   DOI   ScienceOn
7 Bestor TH (2000): The DNA methyltransferases of mammals. Hum Mol Genet 9:2395-2402   DOI   ScienceOn
8 Bird A (2002): DNA methylation patterns and epigenetic memory. Gene Dev 16:6-21   DOI   ScienceOn
9 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   DOI   ScienceOn
10 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
11 Cao H, Stamatoyannopoulos G, Jung M (2004): Induction of human gamma globin gene expression by histone deacetylase inhibitors. Blood 103:701-709   DOI   ScienceOn
12 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   DOI   ScienceOn
13 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   DOI   ScienceOn
14 Stamatoyannopoulos G (2005): Control of globin gene expression during development and erythroid differentiation. Exp Hematol 33:259-271   DOI   ScienceOn
15 Yin H, Blanchard KL (2000): DNA methylation represses the expression of the human erythropoietin gene by two different mechanisms. Blood 95:111-119
16 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   DOI   ScienceOn
17 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   DOI   ScienceOn
18 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
19 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   DOI   ScienceOn
20 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   DOI   ScienceOn
21 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   DOI   ScienceOn
22 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   DOI   ScienceOn