Akt1 players promote PMA U937 cell line differentiation into macrophage-like cells

Abstract

Purpose Monocytes are a leukocytes’ subset that plays an important role in immunity. Protein kinase B (AKT) is involved in monocytes' survival, proliferation and differentiation. Using phorbol 12-myristate 13-acetate (PMA) as an inducer for cell line U937 differentiation into macrophage-like cells may be used as a model for cancer cell therapy or other biomedical research studies. The authors investigated the Akt1 signaling pathway's involvement with PMA as a differentiating agent and survival in the U937 cell line. Design/methodology/approach PMA was utilized to stimulate the differentiation of the U937 cell line into macrophage-like cells at a concentration of 10 nM. Akt1-phosphorylated Serine 473, Bad-phosphorylated Serine 136 and Caspase9-phosphorylated Serine 196 were tested by flow cytometry for the involvement of the Akt1 signaling pathway during differentiation in addition to the expression of CD14, CD206 and CD83. DNA cell cycle variation analysis was done using PI staining and cell viability and apoptosis detection using Annexin V and PI flow cytometry. Findings There was a decrease in phosphorylated Akt1 and Bad activation and an increase in Caspase9 activation, with an increase in surface markers CD14, CD206 and CD83 acquired by PMA-differentiated cells. DNA cell cycle analysis revealed cell accumulation in the G2/M phase and fewer cells in the S phase of PMA-induced U937. Apoptosis induction for Ly294002 or Wortmannin-inhibited cells and part of PMA-induced cells were detected. Originality/value These results may be used to create a model for biomedical research studies and advance the understanding of the mechanism involving differentiation of the U937 cell line.