Tight association of autophagy and cell cycle in leukemia cells

Abstract

Background Autophagy plays an essential role in maintaining cellular homeostasis and in the response to cellular stress. Autophagy is also involved in cell cycle progression, yet the relationship between these processes is not clearly defined. Results In exploring this relationship, we observed that the inhibition of autophagy impaired the G2/M phase-arresting activity of etoposide but enhanced the G1 phase-arresting activity of palbociclib. We further investigated the connection of basal autophagy and cell cycle by utilizing the autophagosome tracer dye Cyto-ID in two ways. First, we established a double-labeling flow-cytometric procedure with Cyto-ID and the DNA probe DRAQ5, permitting the cell cycle phase-specific determination of autophagy in live cells. This approach demonstrated that different cell cycle phases were associated with different autophagy levels: G1-phase cells had the lowest level, and G2/M-phase cells had the highest one. Second, we developed a flow-cytometric cell-sorting procedure based on Cyto-ID that separates cell populations into fractions with low, medium, and high autophagy. Cell cycle analysis of Cyto-ID-sorted cells confirmed that the high-autophagy fraction contained a much higher percentage of G2/M-phase cells than the low-autophagy fraction. In addition, Cyto-ID-based cell sorting also proved to be useful for assessing other autophagy-related processes: extracellular flux analysis revealed metabolic differences between the cell populations, with higher autophagy being associated with higher respiration, higher mitochondrial ATP production, and higher glycolysis. Conclusion This work provides clear evidence of high autophagy in G2/M-phase cells by establishing a novel cell sorting technique based on Cyto-ID.

A novel prognostic prediction model of cuprotosis-related genes signature in hepatocellular carcinoma

“…Moreover, TIGHT regulated TWIST1and promoted vasculature remodeling in bladder cancer ( Liu et al, 2022 ). It also affected autophagy in leukemia and esophageal squamous cell carcinoma ( Gschwind et al, 2022 ; Huang et al, 2023 ). LAG3 was identified as an important therapeutic target in pancreatic cancer, liver, brain, breast cancer and melanoma ( Gulhati et al, 2023 ; Huuhtanen et al, 2023 ; Ulase et al, 2023 ; Zou et al, 2023 ).…”