SDF-1 Gene Expression and Methylation Changes after Isoprenaline Treatment in Human Mesenchymal Stem Cells In vitro

Introduction: A pathway critical to the homing and retention of hematopoietic stem cells (HSCs) within the bone marrow (BM) is the CXCR4/CXCL12. The chemokine stromal cell–derived factor-1 (SDF-1) is highly expressed in human bone marrow endothelium, reticular cells, endosteal osteoblasts and mesenchymal stem cells (MSCs). Neurotransmitters serve as direct chemo attractants to HSPCs and treatment with norepinephrine results in HSC mobilization. In this study, the effect of Isoprenaline as a β-adrenergic agonist on SDF-1 expression and gene promoter methylation status, as a regulatory mechanism of expression gene in human bone marrow MSC was evaluated for the first time. Materials and Methods: MSCs were isolated from human BM and cultured in DMEM plus 15% FBS until passage 3. Then the cells were treated in the presence of100 μM Isoprenaline for 12 and 48 hours. RNA was extracted using Trizol and cDNA synthesized. After setting up RT-PCR for SDF-1, the quantization was performed using qRT-PCR method by ABI Step One device. SDF-1 gene methylation also was assessed using Methylation Specific PCR (MSP) and was analyzed with Totallab Software. Results: The expression level of SDF-1 in 12 and 48 hours post treatment was 7.7 and 0.7 fold in comparison with control sample (P<0.05). SDF-1 gene promoter methylation was increased in 12 and 48 hours after treatment with Isoprenaline. Discussion: The result of this study clarifies the important role of increased level of SDF-1 gene promoter methylation and subsequent reduced level of SDF-1 expression as a key event in stem cell mobilization by regulatory function of nervous system.

کلیدواژه ها:

Mesenchymal stem cells ، SDF-1 ، Methylation ، Isoprenaline