Differential platelet-associated factor gene expression by a panel of human cell lines with megakaryoblastic properties after treatment with phorbol myristate acetate.
Four human megakaryocytoid cell lines, namely MOLM-1, 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.
Micallef
M
M
Fujisaki Institute, Hayashibara Biochemical Laboratories Inc., Okayama, Japan.
Matsuo
Y
Y
Takayama
K
K
Ariyasu
T
T
Otani
T
T
Minowada
J
J
D001483:Base Sequence; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D007124:Immunoenzyme Techniques; D008533:Megakaryocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D010980:Platelet Membrane Glycoproteins / Q000096:biosynthesis; D010982:Platelet-Derived Growth Factor / Q000096:biosynthesis*; D016133:Polymerase Chain Reaction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D019699:Thrombospondins; D013939:Thymidine Phosphorylase / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured; D014841:von Willebrand Factor / Q000096:biosynthesis
D008562:Membrane Glycoproteins; D010980:Platelet Membrane Glycoproteins; D010982:Platelet-Derived Growth Factor; D019699:Thrombospondins; C075278:platelet-derived growth factor A; D014841:von Willebrand Factor; D013939:Thymidine Phosphorylase; D013755:Tetradecanoylphorbol Acetate