Effects Of The Acetoacetate Extract Of Vitex Negundo Seed On The Growth And Apoptosis In Human Acute Myeloid Leukemia HL-60 Cell Line Cultured In Vitro

Objective: To investigate the Effects of the acetoacetate extract of Vitex negundo seed (EVn-50)on the growth and apoptosis of human acute myeloid leukemia HL-60 cells line cultured in vitro.Methods: Human acute myeloid leukemia HL-60 cells and peripheral blood mononuclear cells(PBMC) were cultured in vitro. The cell viability was determinated by MTT assay. The cell growth of HL-60 cells was detected by cell counting. The formation of colony of HL-60 cells was examined by colony formation in soft agar. The apoptosis rate and cell cycle of HL-60 cells was analyzed by flow cytometry with PI staining. The morphous of apoptosis in HL-60 cells was observed by fluorescence microscope after AO/EB staining.Results: 5 species of the extracts from Vitex negundo seed inhibited viability of HL-60 cells, in a concentration-dependent manner. EVn-50 is the strongest of those. IC50 value of that is 2.3μg/mL, which similar to arabinosylcytosin(Ara-C, IC50 is 2.5μg/mL). EVn-50 has little effective that of PBMC, and its IC50 is 25.2μg/mL. EVn-50 possess the relative selectivity for cytotoxicity to HL-60 cells, its selective index is 10.8(25.2/2.3), which is higher than Ara-C(the selective index is 1.7). EVn-50 can efficaciously inhibit the growth and colony formation of HL-60 cells, in a concentration-dependent manner, and has synergism in combination with Ara-C. EVn-50 signifacently induce apoptosis of HL-60 cells, in a concentration-dependent manner, and enhance Ara-C promoting apoptotic effect. Conclusion:1. EVn-50 possess the relative selectivity for cytotoxicity to HL-60 cell line.2. EVn-50 can efficaciously inhibit the growth and colony formation of HL-60 cells, in a concentration-dependent manner.3. EVn-50 signifacently induce apoptosis of HL-60 cells, in a concentration-dependent manner.4 EVn-50 can potentiate Ara-C inhibiting growth and induced apoptosis of HL-60 cells.