Effect Of Artemether On Leukemia Cells In Vitro

Objective:This study was designed to primarily investigate the effects of Artemether on the proliferation,cell cycle,and apoptotic parameters of chronic myelogenous leukemia(CML)-derived K562 cells and mouse leukemic cells L615.To study the effect of Artemether on vascular endothelial growth factor(VEGF)expression in K562 cells and L615 cells and assess the effect of Artemether treatment on endothelial cells proliferation and migration induced by K562 cells and L615 cells in vitro.Methods:MTT(Methy thiazolyl tetrazolium)assay was used to evaluate the inhibitory effects of Artemether treatment on chronic myelogenous leukemia (CML)-derived K562 cells,human umbilical vein endothelial cells(HUV-EC), mouse leukemic cells L615 and mouse pancreatic islet endothelial cells(MS1)in different concentration.FCM(fluorescence flow cytometry)DNA assays and TUNEL (terminal deoxynuxleotidel transferase mediated uridine nucleotide and labeling) assays were applied to detect the changes of apoptotic rate at late apoptosis process and alteration of cell cycle phase.The level of VEGF in conditioned media(CM)of K562 cells and L615 cells pretreated with Artemether was detected by enzyme-linked immunosorbent assay(ELISA)and the form of VEGF mRNA was examined by RT-PCR.MTT assay and scratching assay was used to evaluate the conditioned medium(CM)stimulating effect on endothelial cell proliferation and migration.Results:Artemether could inhibit the proliferation of chronic myelogenous leukemia (CML)-derived K562 cells and mouse leukemic cells L615 and the inhibition depended on the exposure time and dose.The IC₅₀value of K562 cells was 32.27±0.15μg/ml,7.48±0.08μg/ml,and 25.00±0.37μg/ml after treated with Artemether for 24h,48h,and 72h respectively.The IC50 value of L615 cells was 38.07±0.39μg/ml, 27.11±0.71μg/ml,and 25.84±0.32μg/ml after treated with Artemether for 24h,48h, and 72h respectively.The analysis of cell cycle indicated that Artemether blocked cells at G₂/M phases and the cells of G₀/G₁ phase reduced.FCM TUNEL assay indicated that apoptotic rate of K562 cells and L615 cells after treated with Artemether were increased compared with the control cells.The VEGF expression both in K562 cells L615 cells and the conditioned medium(CM)of K562 cells L615 cells pretreated with.Artemether was decrease.The proliferation and migration of human umbilical vein endothelial cells(HUV-EC)and mouse pancreatic islet endothelial cells(MS1)treated with conditioned medium(CM)were decreased compared with the control cells.Conlusion:Artemether could inhibit the proliferation of chronic myelogenous leukemia(CML)-derived K562 cells and mouse leukemic cells L615 and induce cells apoptosis in vitro.Artemether could effectively down-regulate the VEGF expression in K562 cells and L615 cells,and inhibit the endothelial cells proliferation and migration induced by K562 cells L615 cells in vitro.