• YAKHAK HOEJI
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• v.47 no.5
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• pp.319-324
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• 2003

This study examined the inhibitory effect of extracts of Trichosanthes kirilpwii sorted according to the parts on the growth of HL-60 cells. The growth of HL-60 leukemia cells was markedly inhibited by the treatment of the 80％ methanol extract of roots (10 $\mu\textrm{g}$/mι), stems (50$\mu\textrm{g}$/mι), pips (10$\mu\textrm{g}$/mι), and gourds (100 $\mu\textrm{g}$/mι), or the ethylacetate fraction of leaves (100 $\mu\textrm{g}$/mι). when the HL-60 cells were treated with the extracts of T. kirilpwii sorted according to the parts, DNA fragmentation and sub-G1 hypodiploid cells were observed. Moreover, T. kirilpwii extracts increased the level of the expression of the active form of caspase-3 and the activation of caspase-3 was demonstrated by the cleavage of poly(ADP-ribose) polymerase, a vital substrate of effector caspase. The results suggest that the inhibitory effect of extracts of T. kirilpwii sorted according to the parts on the growth of HL-60 cells seems to arise from the induction of apoptosis.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.04a
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• pp.39-55
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• 2003

1. Extracts of the leaves of Eurya emarginata markedly inhibited the growth of leukemia cells such as HL-60, KG-1, U937, K562, and Jurkat. 2. When HL-60 cells were treated with the extract, DNA fragmentation, morphological changes and sub-G1 hypodiploid cells were observed. The expressions of c-myc mRNA were also dramatically decreased. 3. Cornoside and Eutigoside were isolated from the leaves of Eurya emarginata. 4. Eutigoside induced apoptosis through down-regulation of Bcl-2 expression and activation of caspases.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.7
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• pp.819-824
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• 2007

The present study describes the cytotoxic effect of Artemisia fukudo extracts. The extract from A. fukudo by 80% ethanol was fractionated with n-hexane, dichloromethane, ethylacetate, and butanol in serial. The cytotoxicity of A. fukudo extracts was examined for its effect on the growth of HL-60 cells by the colorimetric 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. In addition, we used the HL-60 cells to see what effects the A. fukudo extracts will have on apoptosis of cancer cells. We checked the cell activity, cell morphologic changes, DNA fragmentation, and DNA content after 24 hr incubation with administering 25 ${\mu}g/mL$ of the A. fukudo extracts. In the treatment of the low concentration of n-hexane and dichloromethane fractions, the survival rate of HL-60 cells is lower than that of the control. The laddering-pattern DNA fragmentation was observed in the treatment with n-hexane and dichloromethane fractions. The DNA content of the cells apoptosis measured as the density of sub-$G_{1}$ hypodiploid cells by flow cytometric analysis. The number of sub-$G_{1}$ hypodiploid cells increased in the treatment with n-hexane and dichloromethane fractions. These fractions obstructed the cell cohesion and caused the blebbing of the cell membrane and fragmentation of the nucleus, both of which are symptoms of apoptosis. These results suggest that A. fukudo has a great potential value as food additives, medicinal supplements for patients with chronic diseases, and preventive measures against cancer.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.1
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• pp.187-192
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• 2006

Herbal medicines have been utilized to treat a variety of diseases, including cancer. Several species of the family rubiaceae have been reported to have antitumor activity. In this study, we report the cytotoxicity and antitumor activity exhibited dy the methanol extracts prepared from Rubia radix (RRME), Uncaria gambir (UGME) and Oldenlandia diffusa (ODME) (family: Rubiaceae) against human promyleloid leukemia cell line, HL-60. The cytotoxicity of RRME (2~20 ${\mu}g/ml$), UGME (20~200 ${\mu}g/ml$) and ODME (20~200 ${\mu}g/ml$) were assessed dy the MTT reduction assay. IC50 values for RRME, UGME and ODME were 11.0, 99.5 and 106.1 ${\mu}g/ml$, respectively. When the HL-60 cells were treated with RRME (10 ${\mu}g/ml$), UGME (120 ${\mu}g/ml$) and ODME (140 ${\mu}g/ml$) for 24 h, several apoptotic characteristics such as DNA fragmentation and morphologic changes were observed. Furthermore, flow cytometric analysis was peformed to determine the percent of apoptotic cells. The poupulation of sub-G1 hypodiploid cells was increased 37.49% in RRME treatment, 12.49% in UGME treatment and 7.21% in ODME treatment compared with untreated control cells (2.64%). To further confirm apoptotic cell death, we assayed caspase-3, -8 and -9 activities in RRME, UGME and ODME-treated cells. After treatment of RRME, UGME and ODME for 12 h, caspase-3, -8 and -9 activities significantly increased.compared to untreated control cells. These results show that RRME, UGME and ODME induced apoptotic cell death in HL-60 cells and may have a possibility of potential antitumor activities.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.6
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• pp.1636-1639
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• 2005

To develop novel anti-leukemic medicine, we have prepared a Korean traditional medicine, named Hang-baek-Tang, which is composed of 8 kinds of anti-leukemic medicinal plants. The water extracts was examined anti-leukemic activity using the human leukemia cell line, HL-60 cells. HL-60 cells showed the growth inhibition and several apoptotic features, including DNA ladders, morphological changes, by treatment of the cells with Hang-Daek-Tang. We have observed that Hang-baek-Tang induced the activation of caspase-3, caspase-8 and caspase-9. Further molecular analysis demonstrated that Hang-baek-Tang induced cleavage of PARP and increase of hypodiploid (Sub-G1) population in flow cytometric analysis. These results indicate that Hang-baek-Tang has been considered to exert anti-leukemic activity through the procaspase-3 activation pathway.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.3
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• pp.772-778
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• 2005

The chloromethyl-2-dihydroxyphosphinyl-6,7-dimethoxy-1,2,3,4-tetrahydro- isoquinoline (CDDT) is a newly synthesized derivative from 1,2,3,4-Tetra- hydroisoquinoline (THIQ). The THIQs include potent cytotoxic agents that display a range of antitumor activities, antimicrobial activity, and other biological properties. In this study, we investigated the effect of CDDT on the cytotoxicity, induction of apoptosis in human promyelocytic leukemia cells (HL-60 cells). CDDT showed a significant cytotoxic activity in HL-60 cells ($IC_{50}$ = approximately $37\;{\mu}g/ml$) at a 24 hr incubation. Treatment of HL-60 cells with CDDT displayed several features of apoptosis, including formation of DNA ladders in agarose gel electrophoresis, morphological changes of HL-60 cells with DAPI stain. Here we observed that CDDT caused activation of caspase-3, caspase-8, and caspase-9. The most efficacious time on the activation of caspases-3 was achieved at 12 hr. Further molecular analysis demonstrated that CDDT led to cleavage of poly(ADP-ribose) polymerase (PARP), increase of hypodiploid (Sub-G1) population in the flow cytometric analysis. In conclusion, these above results indicate that CDDT dramatically suppresses HL-60 cell growth by activation of caspase-3 with caspase-8, -9 activity. These data may support a pivotal mechanism for the use of CDDT in the prevention and treatment of leukemia.

• Fisheries and Aquatic Sciences
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• v.13 no.1
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• pp.26-35
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• 2010

In our previous study, we preliminarily demonstrated that Celluclast and Neutrase extracts exhibited the strongest $H_2O_2$-scavenging activities among five carbohydrases (Viscozyme, Celluclast, Termamyl, Ultraflo and AMG) and five proteases (Kojizyme, Alcalse, Flavourzyme Protamex and Neutrase) extracts. Thus, Celluclast and Neutrase extracts were selected for use in further experiments and were separated into four different molecular weight fractions (<5, 5-10, 10-30 and >30 kDa). Among them, the 5-10 kDa fraction showed the highest $H_2O_2$-scavenging activity. The 5-10 kDa fraction also strongly enhanced cell viability against $H_2O_2$-induced oxidative damage. Furthermore, the fraction reduced the proportion of apoptotic cells induced by $H_2O_2$, as demonstrated by decreased sub-G1 hypodiploid cells and decreased apoptotic body formation by flow cytometry. These results indicated that the 5-10 kDa fraction of the Celluclast and Neutrase extracts from S. coreanum exhibited strong antioxidant activity over $H_2O_2$-mediated cell damage in vitro.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.5
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• pp.1370-1374
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• 2005

The Tosyl-JM3 (TJM3) is a modified compound from one of 1,2,3,4-Tetra- hydroisoquinoline (THIQ) derivatives. The THIQs include potent cytotoxic agents that display a range of anti-tumor activities, antimicrobial activity, and other biological properties. In this study, we investigated the effect of TJM3 on the cytotoxicity, induction of apoptosis in human promyelocytic leukemia cells (HL-60 cells). TJM3 showed a significant cytotoxic activity in HL-60 cells (IC50 = approximately $60{\mu}g/m{\ell}$) after a 24 hr incubation. Treatment of HL-60 cells with TJM3 exhibited several features of apoptosis, including formation of DNA ladders in agarose gel electrophoresis, morphological changes of HL-60 cells with DAPI stain. Here we observed that TJM3 caused a decrease of procaspase-3 protein. Further molecular analysis demonstrated that TJM3 led to cleavage of poly(ADP-ribose) polymerase (PARP) by western blot and increase of hypodiploid (Sub-G1) population in the flow cytometric analysis. In conclusion, these above results indicate that TJM3 dramatically suppresses HL-60 cell growth and induces apoptosis. These data may support a possibility for the use of TJM3 in the prevention and treatment of leukemia.

• The Korea Journal of Herbology
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• v.28 no.4
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• pp.33-40
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• 2013

Objectives : To study the apoptosis effects of fermented Undaria pinnatifida extracts(FUP) against HCT-15 colon cancer cells. Method : By measuring cell proliferation, DNA fragmentation, cell cycle, morphology, and western blot from FUP, the study investigated the effects of the extractions had upon the HCT-15 colon cancer cells, and concluded that the inhibiting effects upon cells were induced by apoptosis. Result : FUP effectively inhibited the growth of HCT-15 colon cancer cells. After analyzing the DNA fragmentation, the study observed a DNA ladder, while examining the cells, and found an increase of sub-G1 hypodiploid cells. On the changes regarding the nucleus of the cells, a condensation of cells and chromatin, as well as an apoptotic body was clearly observed. By observing through western blot from FUP, the study found a decreased level of Bcl-2 from HCT-15 colon cancer cells, but the increased level of Bax and cleaved caspase-3, which as a result induced apoptosis, inhibiting the growth of HCT-15 colon cancer cells. FUP increased the natural death of HCT-15 colon cancer cells by the induction of apoptosis. FUP seemed to have no suppressing effect upon HL-60/MX2 cells. However, compared to the fucoidan, the study was able to clearly observe morphological changes of HCT-15 cells apoptosis, in a 1/2 concentration. Conclusion : FUP had antiproliferative effects on different kinds of cancer cells, while proving especially efficacious against colon cancer cells.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.81-81
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• 2003

The present study was taken to examine the inhibitory effect of extracts of Scytosiphon lomentaria, a marine alga growing in Jeju Island, on the growth of cancer cells and to develop an anti-cancer agent using components of S. lomemtaria. The effect was observed by the measurement of metabolic activity using colorimetric 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. In results, crude extract of this alga markedly inhibited the growth of leukemia cell lines such as HL-60 and KG-1, but could scarcely inhibit the growth of normal cells (HEL299) and adenocarcinoma cells (SNU-16 and HCT-I5). When HL-60 cells were treated with the extract, DNA fragmentation and the increase of proportion of sub-G1 hypodiploid cells were observed. Therefore, the inhibitory effect of S. lomemtaria on the growth of HL-60 cells seems to arise from the induction of apoptosis. In order to understand the mechanism of apoptosis inducton by S. lomemtaria, we examined the changes of Bcl-2 and Bax expression. The extract reduced Bcl-2, an anti-apoptotic protein, but increased Bax, a pro-apoptotic protein in a dose-dependent manner. When we examined the activation of caspase-3, an effector of apoptosis, the expression of active form(19 kDa) of caspase-3 was increased and the increase of their activities was demonstrated by the cleavage of poly(ADP-ribose)polymerase, a substrate of caspase-3, to 85 kDa. The results indicate that extract of S. lomentaria induces the apoptosis of HL-60 cells via the down-regulation of Bc1-2 and the activation of caspases.