Elucidating the role of ZRF1 in monocyte-to-macrophage differentiation, cell proliferation and cell cycle in THP-1 cells

Abstract

ZRF1 (Zuotin-related factor 1) is a versatile protein engaged in protein folding, gene regulation, cellular differentiation, DNA damage response, and immune system and cancer development regulation. This study investigates the role of ZRF1 in monocyte-to-macrophage transformation, and its effects on cell proliferation and the cell cycle. We generated ZRF1-depleted THP-1 cells and induced macrophage differentiation using phorbol 12-myristate 13-acetate (PMA). Differentiation was assessed via microscopy and flow cytometry, while cell proliferation was quantified with the [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] (MTS) assay, and the cell cycle was analyzed through flow cytometry using propidium iodide staining. ZRF1-depleted THP-1 cells exhibited notable morphological changes. Flow cytometry post-PMA treatment indicated these cells were smaller and less granular than controls. Proliferation rates of ZRF1-depleted monocytes and macrophages were significantly higher than controls, particularly over longer durations. Cell cycle analysis showed ZRF1 depletion notably affected the G0-G1 phase, highlighting its significant role in macrophage differentiation. The findings provide important insights into ZRF1’s role in monocyte-to-macrophage differentiation and its impact on cell proliferation and the cell cycle. This research not only supports existing knowledge about ZRF1 but also enhances our understanding of its multifaceted roles in cellular processes.