Abstract
Deoxyshikonin was reported to exhibit an anti-tumor effect in colorectal cancer. However, no studies are available to illustrate the effect of deoxyshikonin on acute myeloid leukemia (AML). The effects of deoxyshikonin on viability, apoptosis, caspase-3/7 activity, and cytochrome (Cyt) C expression were evaluated by Cell Counting Kit-8 assay, flow cytometry analysis, caspase-3/7 activity assay, and western blot analysis, respectively. Glucose consumption and lactate production were measured to determine the glycolysis level. The expression of pyruvate kinase M2 (PKM2) was detected by quantitative real-time polymerase chain reaction and western blot analysis. The results showed that deoxyshikonin inhibited cell viability and increased the apoptotic rate, the caspase-3/7 activity, and the Cyt C protein level in AML cells in a dose-dependent manner. Additionally, deoxyshikonin concentration-dependently decreased glucose consumption, lactate production, and PKM2 expression in AML cells. Deoxyshikonin inactivated the protein kinase B (Akt)/mammalian target of the rapamycin (mTOR) pathway. The activation of the Akt/mTOR pathway reversed the effects of deoxyshikonin on viability, apoptosis, and glycolysis in AML cells. In conclusion, deoxyshikonin dampened the viability and the glycolysis of AML cells by suppressing PKM2 via inactivation of the Akt/mTOR signaling.
Highlights
Acute myeloid leukemia (AML), the most prevalent form of acute leukemia in adults, is an aggressive malignancy derived from hemopoietic progenitor cells and with poor survival rate and frequent relapse, posing a threat to the health and life of affected patients [1]
These results suggested that deoxyshikonin facilitated the apoptosis of acute myeloid leukemia (AML) cells
740Y-P resisted the deoxyshikonin-induced decrease of pyruvate kinase M2 (PKM2) protein level (Figures 8D,E) and PKM2 activity (Figure 8F) in THP-1 and HL60 cells. These results suggested that the activation of the Akt/mammalian target of the rapamycin (mTOR) pathway reversed the effects of deoxyshikonin on glycolysis and PKM2 expression in AML cells
Summary
Acute myeloid leukemia (AML), the most prevalent form of acute leukemia in adults, is an aggressive malignancy derived from hemopoietic progenitor cells and with poor survival rate and frequent relapse, posing a threat to the health and life of affected patients [1]. We hypothesized that deoxyshikonin inhibited viability and glycolysis, suppressing pyruvate kinase M2 via the Akt/mTOR pathway in acute myeloid leukemia cells. We assessed the effects and the underlying mechanisms of deoxyshikonin on viability, apoptosis, glycolysis, and PKM2 expression in AML cells. These results revealed that deoxyshikonin treatment inhibited viability, induced apoptosis, MATERIALS AND METHODS. Following the treatments as aforementioned, Apo-ONE Homogeneous Caspase-Glo 3/7 Assay kit (Promega, Madison, WI, USA) was adopted to measure the caspase-3/7 activity of THP-1 and HL60 cells, referring to the manufacturer’s description. RNAiso Plus (TaKaRa, Dalian, China) was used to extract total RNA from treated THP-1 and HL60 cells, and the concentration of extracted RNA was measured using a NanoDrop 2000 spectrophotometer (Thermo Fisher Scientific).
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