Differential platelet-associated factor gene expression by a panel of human cell lines with megakaryoblastic properties after treatment with phorbol myristate acetate
✍ Scribed by Dr. Mark Micallef; Yoshinobu Matsuo; Kazue Takayama; Toshio Ariyasu; Takeshi Otani; Jun Minowada
- Publisher
- John Wiley and Sons
- Year
- 1994
- Tongue
- English
- Weight
- 820 KB
- Volume
- 12
- Category
- Article
- ISSN
- 0278-0232
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✦ Synopsis
Four human megakaryocytoid cell lines, namely MOLM-I, MOLM-7, MEG-01 and HEL, were treated with phorbol 12-myristate 13-acetate (PMA) and expression of the platelet-derived growth factor (PDGF) A chain, von Willebrand factor (vWF) and endothelial cell growth factor (ECGF) genes was examined by the reverse transcriptase-polymerase chain reaction (RT-PCR) using specific primers. The gene for PDGF A chain is constitutively weakly expressed by MEG-01 cells and strong expression is induced in MEG-01. MOLM-1 and MOLM-7 but not in HEL cells after treatment with PMA for three days. All four cell lines express the vWF gene both constitutively and after exposure to PMA. None of the cell lines constitutively express the gene for ECGF but MEG-01 cells can be induced to do so after treatment with the phorbol diester. Immunohistochemical staining after exposure to PMA showed that the expression of the platelet-associated markers CD41 and CD61 was enhanced in all cell lines indicating possible differentiation along the megakaryocyte lineage. Our results illustrate differential platelet-associated factor gene expression in different megakaryoblastic cell populations in response to treatment with PMA, and suggest that expressions of the PDGF A chain gene and the ECGF gene may be good markers for megakaryocyte maturation.
KEY WORDS Megakaryocytes PMA Gene expression lNTRODUCTION Megakaryocytopoiesis and platelet production is an area of study in hematology that remains poorly understood. According to several studies on megakaryocyte proliferation and differentiation, a number of known cytokines such as interleukins 3, 6 and 11, interferon gamma, erythropoietin and leukemia inhibitory factor act at various stages of megakaryocytopoiesis, alone or in c~mbination.'-~ Other investigators have reported the partial characterization of a novel cytokine, isolated from the urine of patients with aplastic anemia, that selectively induces megakaryocyte colony f ~r m a t i o n . ~
The main difficulties encountered in the study of megakaryocyte differentiation arise because of the limited number of megakaryocytes that can be harvested from normal human bone marrow5 as well as the lack of suitable markers for differentiation." To overcome these