Rapamycin induces megakaryocytic differentiation through increasing autophagy in Dami cells

: Autophagy is a conserved cellular process that involves the degradation of cytoplasmic components in eukaryotic cells. However, the correlation between autophagy and megakaryocyte development is unclear. This study aims to explore the role of autophagy in megakaryocyte differentiation. To test our hypothesis, we used the Dami cell line in-vitro experiments. Rapamycin and Bafilomycin A1 were used to stimulate Dami cells. CD41 expression and apoptosis were analysed by flow cytometry. Autophagy-related proteins were detected by Western blotting. 12-O-Tetradecanoylphorbol 13-acetate-treated Dami cells can simulate endomitosis of megakaryocytes in vitro. Rapamycin-induced autophagic cell death was verified by LC3-II conversion upregulation. Meanwhile, Bafilomycin A1 blocked endomitosis and autophagy of Dami cells. Our results provide evidence that autophagy is involved in megakaryocyte endomitosis and platelet development. Rapamycin inhibited cell viability and induced multiple cellular events, including apoptosis, autophagic cell death, and megakaryocytic differentiation, in human Dami cells. Upregulated autophagy triggered by rapamycin can promote the differentiation of Dami cells, while endomitosis is accompanied by enhanced autophagy.