[Alantolactone Inhibits AML Cell Proliferation by Enhancing Reactive Oxygen Species and Downregulating the PI3K/Akt Signaling Pathway].
To investigate the effects of alantolactone (AL) on the proliferation and apoptosis of acute myeloid leukemia (AML) cells and its underlying molecular mechanisms.
AML cell lines SHI-1 and THP-1 were treated with various concentrations of AL. Cell proliferation capacity was assessed using CCK-8 assay, and cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Intracellular reactive oxygen species (ROS) levels were measured using DCFH-DA fluorescent probe, while the mitochondrial membrane potential (MMP) was evaluated using tetramethylrhodamine ethyl ester (TMRE). Western blot analysis was performed to detect the expression of cell cycle regulatory proteins (CDK4, CDK6), apoptosis-related proteins (cleaved caspase-3, Bax, Bcl-XL, PARP), and proteins involved in the PI3K/Akt signaling pathway.
AL significantly inhibited the proliferation of SHI-1 and THP-1 cells in a concentration-dependent (r SHI-1=-0.942 2, r THP-1=-0.882 6) and time-dependent (r SHI-1=-0.979 3, r THP-1=-0.940 0) manner. Meanwhile, AL treatment significantly increased the proportion of cells in G0/G1 phase and apoptosis rate of both SHI-1 and THP-1 cells (P < 0.05), and downregulated the expression of CDK4 and CDK6 (P < 0.05). Additionally, AL significantly upregulated Bax protein expression, downregulated Bcl-XL protein expression, activated caspase-3 and cleaved PARP, thereby promoting cell apoptosis (all P < 0.05). Furthermore, AL treatment significantly elevated intracellular ROS levels and decreased MMP (P < 0.05). Further studies demonstrated that AL could induce programmed cell death in AML cells by inhibiting the activation of the PI3K/Akt signaling pathway.
AL may exert anti-leukemic effects by suppressing AML cell proliferation and inducing apoptosis through multiple mechanisms, including inhibition of PI3K/Akt signaling pathway activation, modulation of Bcl-2 family protein expression, and induction of intracellular oxidative stress.
AML cell lines SHI-1 and THP-1 were treated with various concentrations of AL. Cell proliferation capacity was assessed using CCK-8 assay, and cell cycle distribution and apoptosis rate were analyzed by flow cytometry. Intracellular reactive oxygen species (ROS) levels were measured using DCFH-DA fluorescent probe, while the mitochondrial membrane potential (MMP) was evaluated using tetramethylrhodamine ethyl ester (TMRE). Western blot analysis was performed to detect the expression of cell cycle regulatory proteins (CDK4, CDK6), apoptosis-related proteins (cleaved caspase-3, Bax, Bcl-XL, PARP), and proteins involved in the PI3K/Akt signaling pathway.
AL significantly inhibited the proliferation of SHI-1 and THP-1 cells in a concentration-dependent (r SHI-1=-0.942 2, r THP-1=-0.882 6) and time-dependent (r SHI-1=-0.979 3, r THP-1=-0.940 0) manner. Meanwhile, AL treatment significantly increased the proportion of cells in G0/G1 phase and apoptosis rate of both SHI-1 and THP-1 cells (P < 0.05), and downregulated the expression of CDK4 and CDK6 (P < 0.05). Additionally, AL significantly upregulated Bax protein expression, downregulated Bcl-XL protein expression, activated caspase-3 and cleaved PARP, thereby promoting cell apoptosis (all P < 0.05). Furthermore, AL treatment significantly elevated intracellular ROS levels and decreased MMP (P < 0.05). Further studies demonstrated that AL could induce programmed cell death in AML cells by inhibiting the activation of the PI3K/Akt signaling pathway.
AL may exert anti-leukemic effects by suppressing AML cell proliferation and inducing apoptosis through multiple mechanisms, including inhibition of PI3K/Akt signaling pathway activation, modulation of Bcl-2 family protein expression, and induction of intracellular oxidative stress.