• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.6
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• pp.1074-1077
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• 2004

The effect of gradient eluted fractions from Et$_2$O extracts of Polyporus umbellatus screlotium was investigated on the viability of leukemia cell lines, K-562, L-1210, HL-60 and U-937 cells. Among those fractions, fraction 2 showed mild cytotoxic effect on L-1210 and HL-60 cells. Fraction 3 showed cytotoxic effect on 4 cell lines, and cytotoxic effect was the most potent on L-1210. The hallmark of apoptosis, DNA fragmentation, also appeared by fraction 3 on L-1210 after 48 hr treatment. Furthermore, this fraction was shown to be able to induce cell death on L-1210 cells by the inhibition of DNA synthesis in [$^3$H]thymidine incorporation test. From these results, P. umbellatus involves a potent chemical component that inhibits the viability of leukemia cell lines, L-1210. Further studies about the components of fraction 3 to function as a apoptosis inducer are necessary.

• 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.

• Biomolecules & Therapeutics
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• v.7 no.1
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• pp.22-28
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• 1999

Induction of apoptosis is considered to be the underlying mechanism that accounts for the efficiency of chemotherapeutic drugs. It has recently been proposed that doxorubicin (DOX) can induce apoptosis in human leukemic cells via the Fas/Fas Ligand (FasL) system. Comparison of Fas and FasL mRNA expression between drug- and anti-Fas antibody(Fas-Ab)- induced apoptosis was analyzed for examining the role of Fas/FasL system in the mediation of drug-induced apoptosis. After HL-60 cells were routinely cultured, MTT assay was performed for cytotoxicity test. Giemsa staining was carried out to monitor the apoptosis morphologically. By semiquantitative RT-PCR analysis, the expression of Fas and FasL at 4, 10, 24 hours was determined after DOX and Fas-Ab treatment. Dose-dependent cytotoxicity was induced by DOX-treatment, while Fas-Ab treatment showed the similar dose-dependent pattern but the cytotoxicity is not reached at LD$_{50}$ at 100 ng/ml concentration of Fas-Ab. In the 10ng/m1 DOX and 10ng/m1 Fas-Ab treated group, typical apoptotic cell morphology was shown such as fragmented nuclei and cell membrane budding in the Giemsa-stained slide. Fas mRNA expression was not changed significantly in the both groups. But, FasL mRNA expression was induced significantly at initial period of apoptosis. In this study, Fas/FasL interaction assumed to be involved in drug-induced apoptosis.s.

• The Korean Journal of Nuclear Medicine
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• v.37 no.5
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• pp.308-316
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• 2003

Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.

• Korean Journal of Food Science and Technology
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• v.46 no.6
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• pp.665-670
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• 2014

We isolated twelve lignans and three terpenoids were isolated from the n-hexane fraction of Schisandra chinensis extract. Using spectroscopic data and comparison with available literature, the following compounds were identified: (1) wuweizisu C, (2) gomisin N, (3) deoxyschisandrin, (4) gomisin A, (5) schisandrin, (6) chamigrenal, (7) schisanlactone D, (8) methylgomisin O, (9) angeloylgomisin O, (10) (-)-gomisin $L_2$, (11) schisandronic acid, (12) (-)-gomisin $L_1$, (13) (+)-gomisin $K_3$, (14) gomisin J, and (15) tigloylgomisin H. Notably, this was the first finding that compound (8) was isolated from this plant. Each compound was evaluated for its in vitro cytotoxic activities toward HL-60 (human leukemia), HeLa (human cervical carcinoma), and MCF-7 (breast cancer) cell lines. Compounds (7), (8), and (9) exhibited strong cytotoxic effects on HL-60 ($IC_{50}$ 7.37, 6.60, and $8.00{\mu}M$, respectively), whereas compound (6) exhibited weak cytotoxicity towards MCF-7 ($IC_{50}$ $30.50{\mu}M$). In addition, compound (8) showed the strongest activity towards HeLa cells ($IC_{50}$ $1.46{\mu}M$).

• Journal of Food Hygiene and Safety
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• v.15 no.3
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• pp.241-247
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• 2000

The incidence of verotoxin-producing Escherichia coli(VTEC) was surveyed in domestic foods including hamburger, raw meats and vegetables from 1997 to 1999. The molecular biological characteristics of the isolates were analyzed. Three VTEC strain were isolated from 1,700 samples. Serotypes of those isolates were 0157 : H7, 026 H4, and 056 : Hl2, respectively. Serotype O26 : H4 produced VT I and VT II, and 055 Hl2 isolate produced VT I, however the 60 MDa plasmid DNA and eae gene were not found from both strains. One 0157 : H7 isolate produced VT II and harbour 60 MDa plasmid DNA, however eae gene was not found in the strain. Although they produced VT, it seemed that the virulence of two strains were relatively weak because of the lack of the eae gene. In addition, the serotype O157 : H7 isolate resistant to ampicillin and streptomycin, while isolates of serotype O26 : H4 and O55 : Hl2 were multi-resistant to antibiotics including ampicillin, carbenicillin , cephalothin, trimethoprim/sulfamethoxazole and tetracycline. Supernatants of cultures of all three isolates were showed cytotoxic effect to vero and HeLa cell

• The Journal of the Korean Society for Microbiology
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• v.35 no.4
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• pp.299-308
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• 2000

Recently developed cDNA microarray or DNA chip technology allows expression monitoring of expression of hundreds and thousands of genes simultaneously and provides a format for identifying genes as well as changes in their activity. In order to search for changes in gene expression after X irradiation in HL60 cells, cDNA microarray technique was done. In this study, expression of 588 human genes (including oncogenes, tumor suppressor genes, cell cycle regulator genes, intracellular signal transduction modulator genes, apoptosis related genes, transcription factor genes, growth factors and receptor genes, cytokine genes, etc) were analyzed. For cDNA microarray analysis mRNAs were extracted from control and 8 Gy-irradiated HL60 cells. As a result the changes in expression of several genes were observed. This alteration of gene expression was confirmed by reverse transcription-polymerase chain reaction. The expression of heat shock 60 KD protein, c-jun, erythroid differentiation factor, CPP32, myeloid cell nuclear differentiation antigen, MAP kinase-activated protein kinase, interleukin-8, monocyte chemotactic peptide 1 and RANTES genes was increased, but the expression of p55CDC gene was decreased after X irradiation.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.72-77
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• 2004

Previously, we synthesized a novel Cyclin-dependent kinase inhibitor, MCS-5A. Also, we investigated the involvement of cell cycle regulatory events during MCS-5A-mediated apoptosis in HL-60(+p16/-p53) cells with up-regulation of p16 protein expression. In contrast, apoptosis was not observed in A549(-p16/+p53) cells. Therefore we propose that $p16^{INK4A}$ is a key enzyme for inducing apoptosis. In the present studies, we have explored the mechanism of $p16^{INK4A}$ -mediated cytotoxicity and the role of p16.sup INK4A/ overexpression in the induction of apoptosis in human tumor cells. The tumor suppressor gene $p16^{INK4A}$ is known as a cyclin-dependent kinase inhibitor (CKI) and cell cycle regulator. We expressed wild type $p16^{INK4A}$ in pcDNA3.1 vector and then transfected into non-small cell lung cancer (NSCLC) cell expressing different statue of p16$^{INK4A}$, p53 gene〔A549(-p16/+p53), H1299(-p16/-p53) and HeLa(+pl6/+p53) cell line〕. TUNEL assay (including propidium iodide staining following transfection of these cell line with pcDNA3.1-pl6) indicate that p16$^{INK4A}$-mediated cytotoxicity was associated with apoptosis. This is supported by studies demonstrating an induction of caspase 3 cleavage due to the transfection of A549, H1299 and HeLa cells with pcDNA3.1-pl6. These results suggest that p16$^{INK4A}$ has a new function of inducing apoptosis which is not related with the function of tumor suppressor gene p53.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1028-1035
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• 2004

Imperatorin, a biologically active furanocoumarin from the roots of Angelica dahurica (Umbelliferae), was mutagenic and induced transformation of mouse fibroblast cell lines, whereas it provided inhibiting effects on mutagenesis and carcinogenesis induced by various carcinogens. Furthermore, it has been suggested that imperatorin may have potential anticarcinogenic effects when administered orally in the diet. In addition to its anticarcinogenic properties, imperatorin has been shown to possess anticancer activities. We investigated the macro scale gene expression analysis on the HL-60 cells treated with imperatorin. Imperatorin (10μM) were used to treat the cells for 6h, 12h, 24h, 48h, and 72h. In a human cDNAchip study of 10,000 genes evaluated 6, 12, 24, 48, 72 hours after treated with imperatorin in HL-60 cells. Hierarchical cluster against the genes which showed expression changes by more than 2 fold. Three hundred eighty six genes were grouped into 6 clusters by a hierarchical clustering algorithm. Pathway analysis using gene microarray pathway prof Her that is a computer application designed to visualize gene expression data on screen representing biological pathways and groupings of genes.