• Parasites, Hosts and Diseases
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• v.26 no.4
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• pp.229-238
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• 1988

In vitro culture of Toxoplasma gondii in HL-60 cells and cell-mediates immunity against Toxoplasma in dimethylsulfoxide(DMSO) -induced HL-60 cells, i.e., differentiation into granulocytes, were pursued. HL-60 calls were treated with various concentrations of DMSO, and 1.3%(v/v) for 3 day incubation was chosen as the optimal condition icy differentiation into granulocytes. The degree of differentiation was assayed in physiological and functional aspects in addition to morphological point. When treated with 1.3% DMSO for 3 days, HL-60 cells did not synthesiar DNA materials beyond background level, and showed active chemotactic response to chemotactic peptide, formal-methionyl-leucyl-phenylalanine(FMLP). Morphologically promyelocytes of high nuclearlcytoplasmic(NIC) ratio changed to granulocytes of relatively low WJC ratio. The relationships between HL-60 cells or DMSO-induced HL-60 cells and Toxoplasma were examined after stain with Giemsa and Buorescent dye (acridine orange). HL-60 cells did not show any sign of torso- plasmacidal activity but showed intracellular proliferation of Texoplasma to form rosette for 72 hr co-culture. In contrast, OMSO-induced HL-60 cells phagocytosed Toxoplasma within 1 hr, and performed a process of intracellular digestion of Toxoplasma thereafter. With the above results, it is suggested that phagosome-Iysosome fusion is one of the critical events for the parasitism by Toxoplasma or for susceptibility of host cells. The in vitro culture system of this study has offered a defined condition to study the protozoan parasite-host cell interactions.

• Korean Journal of Veterinary Research
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• v.59 no.4
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• pp.195-199
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• 2019

Disulfiram (DSF) is a member of the dithiocarbamate family that can bind copper. Recent studies have shown that DSF has anti-cancer activities, but the mechanism has not been clarified. Therefore, it is important to study the action mechanism of DSF to maximize its anticancer effects. A human leukemia cell line, HL-60, was used in this study. HL-60 cells were treated with DSF and the cellular metabolic activity was measured. DSF increased the cell death of HL-60 cells in annexin V-fluorescein isothiocyanate/propidium iodide staining analysis. In addition, DSF decreased the mitochondrial membrane potential (MMP) of the HL-60 cells. The cytotoxicity of DSF on HL-60 cells was observed at 0.4 μM. Interestingly, the reduction of MMP by DSF was recovered by N-acetyl-L-cysteine, an inhibitor of reactive oxygen species (ROS) production. This suggests that the decrease in MMP by DSF is closely related to the production of ROS in HL-60 cells, which indicates the relationship between the apoptosis of HL-60 cells by DSF and the role of the mitochondria. This study provides clinicians and researchers with valuable information regarding the anti-cancer activity of DSF in terms of the action mechanism.

• Korean Journal of Veterinary Research
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• v.60 no.2
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• pp.79-83
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• 2020

Fenbendazole (FBZ) is a benzimidazole anthelmintic that has been widely used in treatments for gastrointestinal parasites including pinworms and roundworms in animals. Recently, some studies demonstrated that FBZ has anti-cancer effects related to disruption of microtubule polymerization. In this study, we investigated whether FBZ has anti-cancer activity in HL-60 cells, a human leukemia cell line, and assessed its relationship with the production of reactive oxygen species (ROS). FBZ treatment at 0.25-1 μM significantly decreased the metabolic activity of HL-60 cells. The mitochondrial membrane potential of FBZ-treated HL-60 cells decreased in a concentration-dependent manner. Apoptosis analysis using annexin V-FITC/propidium iodide staining demonstrated that 1 μM FBZ increased the percentages of cells in apoptosis and necrosis. In addition, Hoechst 33342 staining showed the presence of broken nuclei in HL-60 cells treated with 0.5 and 1 μM FBZ. To investigate the anti-cancer mechanism of FBZ, HL-60 cells were treated with FBZ in the absence or presence of N-acetyl cysteine (NAC), an inhibitor of ROS production. NAC significantly recovered the decreased metabolic activity of HL-60 induced by 0.5 and 1 μM FBZ treatments. This study provides evidence that FBZ has anti-cancer activity in HL-60 cells provided, in part, via ROS production.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.66-79
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• 2007

Objectives : This study was carried out to evaluate anti-tummor effect about apoptosis of Cheongyulsodok-Eum (CSE) Results : 1. Anti-tumor(HL-60 cells) effects of CSE water extracts(Exts) were more effective in high density.($IC_{50:}:572$ ${\mu}g/ml$) 2. The generation of $O_2\;^-$ in HL-60 cells were according to the concentration of CSE water Exts, specially more effective on 100 ${\mu}g/ml$ and 1000 ${\mu}g/ml$ concentration. 3. The SOD activities in HL-60 cells were in proportion as cytotoxicity against HL-60 cells of CSE water Exts. 4. The GPx activities in HL-60 cells were in proportion as cytotoxicity against HL-60 cells of CSE water Exts(more effective on 100 ${\mu}g/ml$ and 1000 ${\mu}g/ml$ concentration), but the catalase activities in HL-60 cells were not effective. 5. DPPH radical scavenging activity of CSE water Exts was effective.(3 ${\mu}g/ml:31.2{\pm}5.2$ %, 10 ${\mu}g/ml:49.6{\pm}7.3$ %, 30 ${\mu}g/ml:35.8{\pm}5.7$ % 100 ${\mu}g/ml:42.3{\pm}6.4$ %) 6. The results of cytotoxicity against HL-60 cells of CSE were as follows. 1) In hexane fraction, the cytotoxicity against HL-60 cells($IC_{50:}:592$ ${\mu}g/ml$) was more effective than against NIH3T3 cells. 2) In ethyl acetate fraction, the cytotoxicity against HL-60 cells was not effective. 3) In butanol fraction, the cytotoxicity against HL-60 cell($IC_{50:}:306$ ${\mu}g/ml$) was more effective than against NIH3T3 cells. 4) In $H_2O$ fraction, the cytotoxicity against HL-60 cells was not effective. Conclusion : These result suggest that CSE has antioxidative effects and anti-tumor effects by apoptosis of free radical($O_2\;^-$) activity, especially butanol and hexane fraction from water extract has more effective in anti-twnor effects.

• Biomedical Science Letters
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• v.6 no.3
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• pp.181-186
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• 2000

Asadisulphide were purified from Ferrula assafoetida by organic solvent extraction and chromatography. Its inhibitory effects on the TPA-induced adherence of HL-60 cells was analyzed. Since ethyl acetate extracts of F. assafoetida has the strongest inhibitory effect on adherence of HL-60 cells, it was re-extracted with ethyl acetate, hexane, and ethyl ether and chromatographed three times to isolate asadisulphide. At the minimum concentration of 2 $\mu\textrm{g}$/ml, asadisulphide inhibited adherence of 98% of HL-60 cells that have been treated with TPA. It also showed anti-cancer effect with no cytotoxity in the ED$_{50}$ value of 20 $\mu\textrm{g}$/ml.

• Biomedical Science Letters
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• v.8 no.1
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• pp.13-20
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• 2002

Asadisulphide were purified from Ferrula assafoetida by organic solvent extraction and chromatography. Since ethyl acetate extracts of F. assafoetida has the strongest inhibitory effects on adherence of HL-60 cells, it was reextracted with ethyl acetate, hexane, and ethyl ether and chromatographed three times to isolate asadisulphide. HL-60 cells were grouped into untreated control, TPA-treated, asadisulphide-teated and TPA＋asadisulphide-treated groups, and structural changes of these cells were observed using light microscope, scanning electron microscope and transmission electron microscope to examine the inhibitory effects of asadisulfide on the TPA-induced adherence of HL-60 cells. Light microscopic observations showed that asadisulphide has inhibitory effects on the cell aggregation, extention of cytoplasmic processes and inhibition of substrate adhesion of HL-60 cells. Using scanning and transmission electron microscope, it was observed that cell surfaces and several ultrastructures of TPA-treated HL-60 cell were different from control group, while there were no remarkable differences between asadisulphide-treated and TPA＋asadisulphide-treated group. These results could suggest that asadisulphide has the inhibitory effects on the TPA-induced structural changes of HL-60 cells.

• Toxicological Research
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• v.17 no.2
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• pp.131-137
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• 2001

Baicalein, a major flavonoid of extract from Scutellaria baicalensis Georgi, has been shown to exhibit antioxidant and anti proliferative effects. In the present study, we investigate the effects of baicalein on viability and induction of apoptosis in human promyelocytic leukemia HL-60 cells. Baicalein was found to induce apoptosis of HL-60 cells in a concentration-dependent and time-dependent manner. When HL-60 cells were exposed to 100 $\mu\textrm{M}$ baicalein for 6h, the viability was decreased remarkably to 27% of control, whereas DNA fragmentation was significantly increased to 64%. Nucleosomal fragmentation of baicalein treated HL-60 cells, a hallmark of apoptosis, was further identified by agarose gel electrophoresis (DNA ladder). Flow cytometric analysis showed that apoptotic cells were increased to 66.6% after treatment with 100 $\mu\textrm{M}$ baicalein for 6 h. Baicalein-induced apoptosis of HL-60 cells was reduced by 1h pretreatment with inhibitor of caspases, z-Asp-$CH_2$-DCB. At 3 and 10 $\mu\textrm{M}$ of z-Asp-$CH_2$-DCB, DNA fragmentation of HL-60 cells induced by baicalein (50 $\mu\textrm{M}$) was 36.8 and 17.1 %, respectively, whereas, that of HL-60 cells treated by baicalein (50 $\mu\textrm{M}$) without pretreatment with inhibitor of caspases was 62.7%. These data suggest that baicalein induces apoptosis in human leukemia HL-60 cells, and that caspase enzymes might be involved in baicalein-induced apoptosis.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.3
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• pp.168-171
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• 1998

Sesaminol in sesame seed was postulated to have antitumor activity. The present study was performed to characterize the role of crude sesaminol extracted from sesame seed (Sesame Crude Sesaminol; SCS) on inhibiting the in vitro growth of human leukemia HL-60 cells. SCS inhibited the growth of human leukemia HL 60 cells in culture and macromolecular synthesis in a dose and time dependent manner. The cytostatic range of SCS concentration was found to be 60 to 100 $\mu\textrm{g}$/ml. SCS concentration greater than 200 $\mu\textrm{g}$/mlwere cytocidal to HL-60 cells. When SCS concentraction was 6 $\mu\textrm{g}$/mland 50 $\mu\textrm{g}$/ml the synthesis of HL-60 cells was inhibited by 35% for DNA, 6% for RNA and 5% for protein and 83% for DNA, 76% for RNA and 60% for protein. Of specific interest was the irreversible effect of SCS in inhibiting DNA synthesis of HL-60 cells. This was evidenced from the fact that, even after washed with PBS three times, preincubated HL-60 cells still showed the inhibited DNA synthesis.

• BMB Reports
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• v.33 no.4
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• pp.308-311
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• 2000

The interrelationship between the differentiation and expression of mRNA for transferrin in the HL-60 leukemia cell line was studied. Transferrin mRNA was expressed in HL-60 leukemia cells and the amount was 50% of that in the positive control cell line, HepG-2 cells. The expression of $T_f$ mRNA in HL-60 cells was not regulated by IL-1, IL-6 and $TNF-{\alpha}$, respectively. The expression of $T_f$ mRNA in the differentiated cells into a granulocyte lineage by DMSO, or all-trans RA, was up-regulated (160-170% of control cells); whereas, the expression was not regulated in the differentiated cells into a macrophage lineage by PMA. These results suggest that the differentiation to a granulocyte lineage of HL-60 leukemia cells appear to be related with the upregulation of transferrin mRNA expression.

• YAKHAK HOEJI
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• v.50 no.6
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• pp.372-380
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• 2006

Farnesol in fruits, vegetables, herbs and leaves acts as bioactive component related with prevention of cancer and psychological malaise. We investigated the cytotoxic effects of farnesol on human leukemic cell, HL-60 cells, by MTT assay using 3- (4,5-Oirnethylthiazol-2-yl) -2,5-diphenyltetrazoliumbromide. Farnesol (0.1${\sim}$50 ${\mu}$g/ml) exhibited cytotoxicities against HL-60 cells in concentration and culture period dependent manner, In the cytotoxic condition induced by farnesol against HL-60 cells, the generation of reactive oxygen species such as O$_2$ and H$_2$O$_2$ were found to be considerably increased. The most prominent augmentations of O$_2$ and H$_2$O$_2$ were over five folds of controls. In an attempt to explore the response of HL-60 cells to the increased O$_2$ and H$_2$O$_2$, superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase activities of HL-60 cells treated with farnesol were measured. SOD and GPx activities were found to be remarkably elevated by addition of farnesol showing the best results of 273% and 167% of controls, respectively: All data suggest that farnesol may have played as an apoptosis inducer in HL-60 cells via production of reactive oxygen species (ROS) and HL-60 cells may have failed to overcome the damage of ROS on account of still defcient ROS scavengers including SOD and GPx.