EP2A promotes human CD34+ cell homing in vitro but not proliferation |
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| Authors: | LIU Fang-jie WANG Ya-qun TANG Jing CHEN Hui-zhen LAI Shu-ping SU Chang LI Juan XU Duo-rong |
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| Institution: | Department of Hematology, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou 510080, China |
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| Abstract: | AIM: To investigate the role of prostaglandin E2 receptor 2 agonist (EP2A) in proliferation and homing of human CD34+ cells. METHODS: Bone marrow fluid and peripheral blood containing stem cells were collected from healthy donors mobilized by granulocyte colony-stimulating factor in our department. Human CD34+ cells were isolated by the method of magnetic-activated cell sorting microbeads. Bone marrow mononuclear cells were isolated by Ficoll-Paque centrifugation, and the bone marrow mesenchymal stem cells (BMMSC) were cultured with L-DMEM. Human CD34+ cells and BMMSC were divided into 4 groups, and treated with PGE2 (as positive control), DMSO (as negative control), EP2A and EP2A+prostaglandin E2 receptor 2 antagonist (EP2AA), respectively. After exposed to the reagents, human CD34+ cell viability was measured by CCK-8 assay, the number of colonies was evaluated by colony-formation assay, the cell cycle distribution was analyzed by flow cytometry, and the protein expression of survivin, β-catenin and CXC chemokine receptor 4 (CXCR4) was detrmined by Western blot. Moreover, the concentration of stromal cell-derived factor-1α (SDF-1α) in the BMMSC was detected by ELISA. RESULTS: The cell viability and the colony number of human CD34+ cells in EP2A group were not higher than those in negative control group. Furthermore, the proportion of human CD34+ cells treated with EP2A in G2/M phase was not elevated compared with negative control group. The protein expression of survivin and β-catenin did not up-regulated in human CD34+ cells exposed to EP2A, but the protein expression of CXCR4 in human CD34+ cells and the concentration of SDF-1α in BMMSC were elevated. CONCLUSION: EP2A promotes human CD34+ cell homing in vitro but not proliferation. |
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| Keywords: | Prostaglandin E2 Prostaglandin E2 receptor 2 agonist Cell proliferation Homing |
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