| Background: Dehydrocostus lactone is an active monomer ingredient extracted from Costustoot,which has been proved to have antibacterial,anti-inflammatory and anti-tumor effects in recent years.Acute myeloid leukemia(AML)is the most common type of acute leukemia in adults,which has a high recurrence rate and mortality.The development of new effective therapeutic drugs is still one of the focuses in the field of acute leukemia.Objective: To investigate the inhibitory effect of dehydrocostus lactone on proliferation of AML cells and its possible molecular mechanism.Methods: Acute myeloid leukemia cell lines U937,MOLM-13,THP-1,and MV4-11 were selected as the research objects in this experiment.CCK-8 assay,trypan blue exclusion assay and methylcellulose clone formation assay were used to detect the inhibitory effect of dehydrocostus lactone on the growth of AML cell lines.Flow cytometry was used to detect the cell cycle distribution and apoptosis of U937 and MOLM-13 cells after treatment with various concentration of dehydrocostus lactone.Western blot was used to detect the expression of apoptosis-related proteins(cleaved-caspase-3,cleaved-PARP,and p-?H2AX),cell cycle-related proteins(CDK4,cyclin D,CDK1,and cyclin B1)and PI3K-AKT-m TOR signaling pathway related proteins(phospho-S6,phospho-AKT)in U937 and MOLM-13 cells after treatment with different concentrations of dehydrocostus lactone.Results: 1.The results showed that dehydrocostus lactone could significantly inhibit the proliferation of U937,MOLM-13,THP-1,and MV4-11 cells after treatment with different concentrations of dehydrocostus lactone for 72 h in a dose-dependent manner.The IC50 values of dehydrocostus lactone against U937,MOLM-13,THP-1,and MV4-11 were around 6.003 ± 0.809 ?M,3.138 ± 0.525 ?M,3.737 ± 0.633 ?M,and6.994 ±0.911 ?M respectively.Moreover,the anti-leukemic effect of dehydrocostus lactone was also evaluated using a colony-formation assay in methylcellulose-based medium.Treatment with dehydrocostus lactone at various concentrations for 10 days was found to significantly inhibit the colony-forming ability of U937 and MOLM-13 cells.U937 and MOLM-13 cells were treated with 0,4,6,and 8 ?M dehydrocostus lactone,and the flow cytometry results demonstrated that dehydrocostus lactone dramatically induced apoptosis.The apoptosis rates of U937 cells were 5.5 ± 1.3%,7.7± 1.8%,16.6 ± 2.4%,21.5 ± 4.3% respectively,and the MOLM-13 cell apoptosis rate was significantly increased from 6.1 ± 1.8 % in the control group to 10.4 ± 2.0%,19.4 ±4.2%,39.5 ± 5.1%.The cell cycle phase distribution was analyzed using flow cytometry.The results showed that dehydrocostus lactone could significantly induce cell cycle arrest at the G0/G1 phase in U937 cells and induce cell cycle arrest at G2/M phase in MOLM-13 cells.Our results showed that dehydrocostus lactone significantly induced cell apoptosis by upregulating cleaved-caspase-3,cleaved-PARP,and p-?H2AX exprssion.We also investigated the expression of the cell cycle relative proteins.The results revealed that the expression of cyclin D and CDK4 was gradually downregulated in U937 cells with increasing concentrations of dehydrocostus lactone,and the protein levels of CDK1 and cyclin B1 were decreased in MOLM-13 cells after treatment with dehydrocostus lactone.Our results revealed that dehydrocostus lactone significantly inhibited the expression of p-AKT and p-S6 proteins,suppressed the proliferation of AML cell lines via inhibition of PI3K-AKT-m TOR signaling pathway.Conclusion:1.Dehydrocostus lactone inhibits the proliferation of AML cell lines U937 and MOLM-13 through cell cycle areest.2.Dehydrocostus lactone can induce apoptosis of U937 and MOLM-13 cells.3.The anti-leukemic effects of dehydrocostus lactone maybe associated with inhibition of the PI3K-AKT-m TOR signaling pathway,which may useful for acute myeloid leukemia. |
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