J Korean Med Sci.  2005 Jun;20(3):409-416. 10.3346/jkms.2005.20.3.409.

Direct and Indirect Effects of Androgens on Survival of Hematopoietic Progenitor Cells In Vitro

• Kim SW
• Hwang JH
• Cheon JM
• Park NS
• Park SE
• Park SJ
• Yun HJ
• Kim S
• Jo DY

• KMID: 1778498
• DOI: http://doi.org/10.3346/jkms.2005.20.3.409

Abstract

Androgens remain a common treatment for certain type of anemia, based upon its myelostimulating effects; however, it has not been established whether androgens affect apoptosis of hematopoietic progenitor cells (HPCs). We investigated the effects of the androgens, such as testosterone, 5beta-dihydrotestosterone (5-DHT), and oxymetholone, on apoptosis of normal hematopoietic progenitor cells in vitro. Androgens did not rescue normal bone marrow (BM) CD34+ cells and colony-forming cells (CFCs), other than mature erythroid CFCs, from apoptosis induced by serum- and growth factor deprivation. Oxymetholone did not affect growth factor-mediated survival of normal CD34+ cells or its inhibition by interferon-gamma (IFN-gamma). In a standard methylcellulose clonogenic assay, low concentrations of oxymetholone and 5-DHT stimulated the clonal growth of colony-forming unit (CFU)-erythroid, but did not affect growth of CFU-granulocyte/macrophage or burst-forming unit-erythroid. Oxymetholone and 5-DHT stimulated the production of stem cell factor in normal bone marrow stromal cells (BMSCs) via transcriptional regulation. In agreement with this, oxymetholone-treated BMSCs better supported the survival of HPCs. These data indicate that survival-enhancing or growth-stimulatory effects of androgens on hematopoietic progenitor cells are minimal and mostly restricted to mature erythroid progenitors, and its myelostimulating effects could be attributed, at least in part, to the stimulation of production of hematopoietic growth factors in BMSCs.

Keyword

Androgens; Erythroid Progenitor Cells; Myeloid Progenitor Cells; Apoptosis

MeSH Terms

Androgens/*pharmacology
Antigens, CD34/analysis
Apoptosis/drug effects
Blotting, Northern
Blotting, Western
Bone Marrow Cells/cytology/drug effects/immunology
Cell Survival/drug effects
Cells, Cultured
Chemokines, CXC/genetics/metabolism
Colony-Forming Units Assay
Cytokines/genetics/pharmacology
Dihydrotestosterone/pharmacology
Dose-Response Relationship, Drug
Flow Cytometry
Gene Expression/drug effects
Hematopoietic Stem Cells/cytology/*drug effects/metabolism
Humans
Oxymetholone/pharmacology
RNA, Messenger/genetics/metabolism
Research Support, Non-U.S. Gov't
Reverse Transcriptase Polymerase Chain Reaction
Testosterone/pharmacology
Time Factors

Figure

• 

  Fig. 1 Effects of androgens on apoptosis of normal bone marrow CD34+ cells induced by growth factor deprivation. BM CD34+ cells were incubated in IMDM for 36 hr in the presence or absence of the indicated hormones (10-5 M), stained with annexin V and propidium iodide, and analyzed on a flow cytometer. (A) Mean percentages and SD of annexin V-positive cells from five independent experiments are shown. *p<0.05 compared with untreated control (IMDM containing 0.1% ethanol) (analyzed by t-test for paired samples). (B) 5-DHT (10-5 M) increased the percentage of annexin-positive cells. The addition of either SDF-1 (100 ng/mL) or SCF (100 ng/mL) in combination with 5-DHT partially relieved the increase in apoptosis induced by 5-DHT, and the addition of both SDF-1 and SCF completely reversed it. A representative result is shown.

• 

  Fig. 2 5-DHT, but not oxymetholone, down-regulates the production of SDF-1 in bone marrow stromal cells. (A) Primary human bone marrow stromal cells (BMSCs) were incubated in serum-free medium X-VIVO with or without (control) the indicated androgens. After a 72-hr incubation, the expression of SDF-1 was analyzed in total cell lysates by Western blotting. (B) 5-DHT down-regulates the expression of SDF-1 mRNA in BMSCs. Primary human BMSCs were incubated in serum-free medium X-VIVO in the presence of 5-DHT (10-5 M). At the indicated time points, the level of SDF-1 mRNA in the cells was analyzed by Northern hybridization. A blot of 28S ribosomal RNA is shown to control for RNA loading.

• 

  Fig. 3 Oxymetholone and 5-DHT modulate the levels of the mRNAs of various cytokines in bone marrow stromal cells (BMSCs). Primary human BMSCs were incubated in serum-free medium X-VIVO for 24 hr with or without (control) the indicated androgens (10-5 M) and then analyzed by RT-PCR.

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