• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.3
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• pp.677-683
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• 2005

The aim of this study was to investigate the apoptotic effect and its mechanism on Radix Aconiti (RA) extract in HL-60 human leukemia cell line. RA extract induced apoptosis as confirmed by discontinuous fragmentation of DNA. To clarify the mechanisms on RA extract-induced apoptosis, we examined the caspase-3, -8 enzyme activity and protein levels including Fas, FasL in HL-60 cells. Treatment with RA extracts resulted in the increase of caspase-3 enzyme activity in a time and dose-dependent manners, which was accompanied by the cleavage of poly-(ADP-ribose) polymerase (PARP). This activation of caspase-3 enzyme resulted from cleavage of procaspase-8, which was followed by increases of FasL, Fas protein expression in RA extracts-treated HL-60 cells. In conclusion, RA extract induced apoptosis of HL-60 human leukemia cell line. This results suggest that the apoptotic mechanisms of RA extract on HL-60 cells involved in FasL, Fas activation, procaspase-8 cleavage, activation of caspase-3 and cleavage of PARP. Collectively, these results suggest that RA may be a valuable agent as a anti-cancer drug.

• 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 ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.99-111
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• 2000

Objectives: Hedyotis diffusa is used to treat cancer in traditional Korea Medicine. So this study was carried out to examine the expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa. Methods: 1. HL-60 cells were treated with various concentrations (from 200 to $50{\mu}g/ml$)of methanol extract and H20 extract ($200{\mu}g/ml$) of hedyotis diffusa. After 48 h later, the cells were tested for viability by MTT assay. 2. The HL-60 cells were treated with $200{\mu}g/ml$ of methanol extract for the indicated periods. The whole cell lysates were prepared and analyzed by western blotting using anti-p53 antibody. 3. The nuclear extract were prepared and analyzed by western blotting using anti-p21 antibody, anti-p27 antibody, anti-cyclin A antibody, anti-cyclin E antibody and anti-CDK2 antibody. Results: 1. The methanol extract of Hedyotis diffusa induced the death of HL-60 cells in a dose dependent manner. 2. The methanol extract of Hedyotis diffusa makedly decreased the level of p21/Cipl and cyclin A in a time dependent manner. 3. The methanol extract of Hedyotis diffusa markedly increased the level of p27/Kipl and cyclin E in a time dependent manner. 4. The methanol extract of Hedyotis diffusa markedly did not affect the level of CDK2. Conclusions: These results provide evidence that expression of cell cycle related genes in HL-60 cells undergoing apoptosis by the methanol extract of Hedyotis diffusa mainly results from decreased level of p21/Cipl and increased level of p27/Kipl of the cell cycle related genes.

• Radiation Oncology Journal
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• v.16 no.4
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• pp.377-388
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• 1998

Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

• Biomedical Science Letters
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• v.19 no.2
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• pp.118-123
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• 2013

Tumor necrosis factor-alpha (TNF-${\alpha}$) is a proinflammatory cytokine that mediates the inflammatory response and immune functions, and modulates the proliferation, differentiation and cell death of cancer cells. The differential functions of TNF-${\alpha}$ in various human cells due to the formation of different stimulating pathway upon the binding of TNF-${\alpha}$ to its receptors. In the present study, we examined the different effects of TNF-${\alpha}$ on the survival and apoptosis between normal granulocytes and human myeloid leukemia HL-60 cells. Although TNF-${\alpha}$ did not affect on the constitutive apoptosis of granulocytes, TNF-${\alpha}$ strongly induced the apoptosis of HL-60 cells in a dose- and a time-dependent manner. TNF-${\alpha}$-induced apoptosis was occurred via the activation of caspase 8, caspase 9 and caspase 3/7 and the induction of ROS production in HL-60 cells. Also, BAY-11-7085, a NF-${\kappa}B$ inhibitor, blocked the TNF-${\alpha}$-induced apoptosis in HL-60 cells. NF-${\kappa}B$ may be involved in TNF-${\alpha}$-induced apoptotic signaling pathway in HL-60 cells. These results suggest that TNF-${\alpha}$ activates apoptotic pathways and its process depends on cell type and many cellular factors. A better understanding of the differential effect of TNF-${\alpha}$ on cell apoptosis and survival may provide important information that can be used to elucidate the specific inhibitory effect of TNF-${\alpha}$ on the cancer dis.

• Toxicological Research
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• v.15 no.1
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• pp.19-25
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• 1999

(+)-Catechin inhibited the growth and induced the differentiation of HL-60 human leukimia cells. The degree of a differentiation by (+)-catechin during the differentiation, the expression assay, To understand the molecular mechanism of (+)-catechin during the differentiation, the expression level of oncogenes was detected by Northern blot analysis. c-Myc mRNA level was reduced after treatment with (+)-Catechin (10-4), however, the expression of c-jun was increased with a concentration dependent manner in HL-60 cells. These results showed that the differentiation and antiproliferation of HL-60 cells against (+)-Catechin was related to the reduction of c-myc and the induction of c-jun expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.4
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• pp.903-907
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• 2005

Many naturally occurring plant extracts are studied for their beneficial effects for health and particularly on cancer. Apoptosis, or programmed cell death, occurs in both normal and pathological conditions, including cancer Dysregulation of apoptosis allows transformed cells to continually and uninhibitedly enter the cell cycle, thus perpetuating the sequence of mutation, genomic instability and, finally, oncogenesis. To investigate the apoptosis-inducing effect of the extract of Fructus Trichosanthis (EFT) on leukemic HL-60 cells and its mechanism, HL-60 cells in vitro in culture medium were given different doses of the extract. The inhibitory rate of cells were measured by microculture tetrazolium assay, cell apoptotic rate was detected by flow cytometry, morphology of cell apoptosis was observed by DAPI fluorescence staining, and the activations of caspases and PARP were detected using Western blotting analysis. The extract could activate the caspase-3 and caspase-8, induce PARP cleavage, inhibit growth of HL-60 cells, and cause apoptosis significantly The suppression was in dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by DAPI fluorescence staining especially. These results will provide strong laboratory evidence of EFT for clinical treatment of acute leukemia.

• The Journal of Traditional Korean Medicine
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• v.15 no.1
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• pp.83-89
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• 2006

Many naturally occurring plant extracts are studied for their beneficial effects for health and particularly on cancer. Apoptosis, or programmed cell death, occurs in both normal and pathological conditions, including cancer. Dysregulation of apoptosis allows transformed cells to continually and uninhibitedly enter the cell cycle, thus perpetuating the sequence of mutation, genomic instability and, finally, oncogenesis. To investigate the apoptosis-Inducing effect of the extract of Fructus Trichosanthis (EFT) on leukemic HL-60 cells and its mechanism, HL-60 cells in vitro in culture medium were given different doses of the extract. The inhibitory rate of cells were measured by microculture tetrazolium assay, cell apoptotic rate was detected by flow cytometry, morphology of cell apoptosis was observed by DAPI fluorescence staining, and the activations of caspases and PARP were detected using Western blotting analysis. The extract could activate the caspase-3 and caspase-8, induce PARP cleavage, inhibit growth of HL-60 cells, and cause apoptosis significantly. The suppression was in dose-dependent manner. Marked morphological changes of cell apoptosis including condensation of chromatin and nuclear fragmentation were observed clearly by DAPI fluorescence staining especially. These results will provide strong laboratory evidence of EFT for clinical treatment of acute leukemia.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.6
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• pp.1190-1196
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• 2002

Selaginella Tamariscina is widely used in the traditional oriental herbal medicine for its anti-inflammatory, anti-cancer effects. The effects of aqueous extracts of Selaginella Tamariscina (ST) on the cell viability and induction of apoptotic cell death were investigated in A549, Raw 264.7, C6-glioma. Jurkat and HL-60 cells. The cell viability after treating with extract of Selaginella Tamariscina was quantified by MTT assay method. The results showed that ST decreased the cell viability in HL-60 and Jurkat cells not in A549, Raw 264.7 and C6-glioma cells. And we also observed the chromatin condensation and DNA fragmentation in HL-60 and Jurkat cells. The enzyme activity of caspase-3, tightly regulated by an apoptosis activating complex, were markedly increased in HL-60 cells treated with the ST by dose-dependent manner. In conclusion, our results suggest that the extract of Selaginella Tamariscina may induce the selective apoptotic cell death in HL-60 and Jurkat cells via activation of caspase-3.

• Archives of Pharmacal Research
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• v.17 no.1
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• pp.45-47
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• 1994

A cytotoxic sesquiterpene against L1210 and Hl-60 cells was isolated from Angelicae Koreanae Radix (bulk-kang-hwal). The component was identified as bisabolangelone by means of chemical and physical methods. The $ED_{50}$ values of it were $1.20{\;}\mu\textrm{g}/ml$ against L1210 cells and $2.30{\;}\mu\textrm{g}/ml$ against HL-60 cells. Bisabolangelone was found in bulk-kang-hwal but not in kang-hwal.