• Archives of Pharmacal Research
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• v.29 no.12
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• pp.1140-1146
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• 2006

Ceramide analogs are potential chemotherapeutic agents. We report that a ceramide analog induces apoptosis in human prostate cancer cells. The ceramide analog induced cell death through an apoptotic mechanism, which was demonstrated by DNA fragmentation, the cleavage of poly ADP ribose polymerase (PARP), and a loss of membrane asymmetry. Treating the cells with ceramide analog resulted in the release of various proapoptotic mitochondrial proteins including cytochrome c and Smac/DIBLO into the cytosol, and a decrease in the mitochondrial membrane potential. In addition, the ceramide analog decreased the phospho-Akt and phospho-Bad levels. The expression of the antiapoptotic Bcl-2 decreased slightly with increasing Bax to Bcl-2 ratio. These results suggest that the ceramide analog induces apoptosis by regulating multiple signaling pathways that involve the mitochondrial pathway.

• Biomolecules & Therapeutics
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• v.20 no.1
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• pp.62-67
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• 2012

TNF-related apoptosis-inducing ligand (TRAIL) is a promising agent for management of cancer because of its selective cytotoxicity to cancer cells. However, some cancer cells have resistance to TRAIL. Accordingly, novel treatment strategies are required to overcome TRAIL resistance. Here, we examined the synergistic apoptotic effect of apigenin in combination with TRAIL in Huh-7 cells. We found that combined treatment of TRAIL and apigenin markedly inhibited Huh-7 cell growth compared to either agent alone by inducing apoptosis. Combined treatment with apigenin and TRAIL induced chromatin condensation and the cleavage of poly (ADP-ribose) polymerase (PARP). In addition, enhanced apoptosis by TRAIL/apigenin combination was quantified by annexin V/PI flow cytometry analysis. Western blot analysis suggested that apigenin sensitizes cells to TRAIL-induced apoptosis by activating both intrinsic and extrinsic apoptotic pathway-related caspases. The augmented apoptotic effect by TRAIL/apigenin combination was accompanied by triggering mitochondria-dependent signaling pathway, as indicated by Bax/Bcl-2 ratio up-regulation. Our results demonstrate that combination of TRAIL and apigenin facilitates apoptosis in Huh-7 cells.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2004.06a
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• pp.297-301
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• 2004

The purpose of the present study was to investigate the anti-proliferative and apoptotic effects of MCS-C2, a novel analogue of toyocamycin and sangivamycin, in human promyelocytic leukemia (HL-60) cells. When treated with MCS-C2, inhibited proliferation associated with cell cycle arrest and apoptotic induction was found in the HL-60 cells in a concentration-dependent and time-dependent manner. This apoptotic induction was associated with the cleavage of Bid and a release of cytochrome c from mitochondria into the cytosol, followed by the activation of caspase-3 and inactivation of poly (ADP-ribose) polymerase (PARP). However, there was no significant change in any other mitochondrial membrane proteins, such as Bcl-2 and Bax. Consequently, the current findings suggest that the mitochondrial pathway was primarily involved in the MCS-C2-induced apoptosis in the human promyelocytic leukemia HL-60 cells.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.95.2-96
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• 2003

Ultraviolet B (UVB) is known to induce apoptosis in human melanocytes. Here we show the cytoprotective effect of sphingosine-1-phosphate (S1P) against UVB-induced apoptosis. We also show that UVB-induced apoptosis of melanocytes is mediated by caspase-3 activation and poly(ADP-ribose) polymerase (PARP) cleavage, and that S1P prevents apoptosis by inhibiting this apoptotic pathway. We further investigated three major subfamilies of mitogen-activated protein (MAP) kinases and the Akt pathway after UVB irradiation. (omitted)

• The Journal of Pediatrics of Korean Medicine
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• v.20 no.1
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• pp.257-272
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• 2006

Objective : In this study, the ability of Oriental medicine Samultanggamibang(SMTG) to induce apoptosis was investigated in B16F10 melanoma cells. Method : Tetrazolium-based colorimetric assay was performed for cytotoxicity test. Several new assays for the basis of biochemical events associated with apoptosis such as DNA fragmentation by a flow cytometry, caspase-3 activation and PARP cleavage by Western blotting should be carried out potentially useful for the basis of biochemical events associated with apoptosis such as a flow cytometry and caspase-3 activation. Results : (1) The number of B16F10 melanoma cells was less than 30 % after exposure to 1 mg/ml SMTG for 48 h. SMTG increased cytotoxicity of B16F10 melanoma cells in a dose- and time-dependent manner. (2) The percentage of apoptotic cells by flow cytometric analysis of the DNA-stained cells increased to 21 % at 24 h and 25 % at 48 h after treatment with 1 mg/ml SMTG. (3) SMTG-induced apoptosis was accompained by the activation of caspase-3 and the specific proteolytic cleavage of poly-ADP-ribose polymerase. (4) SMTG induces the activation of caspase-3 and the specific proteolytic cleavage of poly-ADP-ribose polymearse and eventually leads to apoptosis through c-Jun NH2-terminal protein kinase (JNK)-dependent manner in B16F10 melanoma cells. Conclusion : SMTG had a strong cytotoxic effect of B16F10 melanoma cells.

• Toxicological Research
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• v.25 no.3
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• pp.113-118
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• 2009

Cell death induced by menadione (vitamin K-3,2-methyl-1,4-naphthoquinone) has been investigated in human promyelocytic leukemia HL-60 cells. Menadione was found to induce both apoptosis and necrosis in HL-60 cells. Low concentration ($1{\sim}$50 ${\mu}$M) of menadione induced apoptotic cell death, which was demonstrated by typical DNA ladder patterns on agarose gel electrophoresis and flow cytometry analysis. In contrast, a high concentration of menadione (100 ${\mu}$M) induced necrotic cell death, which was demonstrated by DNA smear pattern in agarose gel electrophoresis. Necrotic cell death was accompanied with a great reduction of cell viability. Menadione activated caspase-3, as evidenced by both increased protease activity and proteolytic cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP) into 85 kDa cleavage product. Caspase-3 activity was maximum at 50 ${\mu}$M of menadione, and very low at 100 ${\mu}$M of menadione. Taken together, our results showed that menadione induced mixed types of cell death, apoptosis at low concentrations and necrosis at high concentrations in HL-60 cells.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.13-24
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• 2006

Purpose : 이 연구는 MCF-7 인간 유방암 세포주에 대한 활락효령단(活絡效靈丹) 추출물의 증식억제효과, 세포독성효과, 세포사 유발효과를 확인하기 위하여 이루어졌다. Methods : MCF-7 인간 유방암 세포주는 Dulbecco's modified Eagle's medium/F12 (DMEM/F12)에 10% fetal bovine serum (FBS)와 항생제를 가하여 만든 배지를 이용하여 배양하였고 MCF-7 세포를 96-well plate에 접종한 후 다양한 농도 (0 ${\sim}$ 2000 g/ml)의 활락효령단(活絡效靈丹)이 든 배지로 처리한 후 72시간 동안 배양하였고 또한 1000g/ml의 활락효령단(活絡效靈丹)이 든 배지로 처리한 후 48, 96, 192 시간동안 배양하여 각각 MTS assay kit로 세포생존율을 측정하였다. 세포독성은 Sulforhodamine B assay 방법을 이용해 측정하였고 세포사 과정에서 MCF-7 세포에서의 caspase 활성화를 측정하기 위해 Western blotting을 수행하여 poly ADP ribose polymerase (PARP)의 절단을 확인하였다. Results : 실험결과 활락효령단(活絡效靈丹) 추출물에 의한 세포성장 및 독성효과는 시간 및 농도에 비례하는 것으로 나타났고 세포고사과정에서 작용하는 caspase의 전기질인 PARP 절단량이 활락효령단(活絡效靈丹) 처리 농도에 비례에 증가하였다. Conclusions : 활락효령단(活絡效靈丹)은 다양한 기전에 의해서 유방암 세포에 대한 억제효과를 가질 수 있는 것으로 인식할 수 있다.

• Archives of Pharmacal Research
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• v.28 no.1
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• pp.68-72
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• 2005

Pseudolaric acid B is a major compound found in the bark of Pseudolarix kaempferi Gordon. In our study, pseudolaric acid B inhibited growth of human melanoma cells, A375-S2 in a time and dose-dependent manner. A375-S2 cells treated with pseudolaric acid B showed typical characteristics of apoptosis including morphologic changes, DNA fragmentation, sub-diploid peak in flow cytometry, cleavage of poly-ADP ribose polymerase (PARP) and degradation of inhibitor of caspase-activated DNase (ICAD). P53 protein expression was upregulated while cells were arrested at the $G_2/M$ phase of the cell cycle. There was a decrease in the expression of anti-apoptotic Bcl-2 and Bcl-xL proteins, whereas pro-apoptotic Bax was increased. The two classical caspase substrates, PARP and ICAD, were both decreased in a time-dependent manner, indicating the activation of downstream caspases.

• YAKHAK HOEJI
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• v.48 no.2
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• pp.153-158
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• 2004

Paecilomyces tenuipes is one of the famous Chinese medicinal entomopathogenic fungi that parasite in the lavae of silkworm. A cytotoxic compound, 4$\beta$-acetoxyscirpendiol (ASD) was isolated from a methanolic extracts of Paecilomyces tenuipes. The ASD compound belongs to scirpenol subfamily of trichothecene mycotoxin. In a continuation of the elucidation of the mechanism of ASD, we report here the evidences of induction of apoptosis by ASD in human Jurkat T cell line. In MTT reduction assay for monitoring cell viability, ASD showed strong toxicity. The 50 percent inhibitory concentrations of ASD against human T lymphoid Jurkat cell was 59.5 ng/$m\ell$. Phosphatidylserine externalization was increased by ASD at 3 and 6 hrs when compared with that of 6 hrs in the cell line showing in a time-dependent manner. When whole lysates of cells treated with ASD were subjected to western blot assay, 113 kDa poly(ADP-ribose) polymerase (PARP) was significantly cleaved to 89 kDa fragment. Time-dependent DNA fragmentation was also observed when Jurkat T cells were treated with ASD at 100 ng/$m\ell$ for 6 hrs and 18 hrs at the ratios of 8.5% and 15.0%, respectively. From these data, Jurkat T lymphocytes treated with ASD from Paecilomyces tenuipes underwent typical cascades of apoptotic cell death.

• Journal of Oriental Neuropsychiatry
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• v.11 no.2
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• pp.1-9
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• 2000

In previous studies, heat shock has been reported to induce the apoptosis or programmed cell death through the activation of caspase-3. 1 investigated the effect of juniper pure essential oil on the heat shock-induced apoptosis in human astrocyte cell line CCF-STTGI. Treatment of the astrocytes with heat shock markedly induced apoptotic cell death. However, pretreatment of the astrocytes with juniper oil ingibited the heat shock-induced apoptosis. To determine whether juniper inhibits the heat shock-induced activation of these apoptotic proteases, activation of CPP32 was assessed by Western blotting. Consistent with flow cytometry. DNA fragmentation and giemsa staining, heat shock-induced activation of CPP32 was blocked by juniper oil. Poly(ADP-ribose) polymerase (PARP), cysteine protease substrates were fragmented as a consequence of apoptosis by heat shock. Juniper oil inhibited the PARP fragmentation. This juniper oil also inhibited the heat shock-induced activation of caspase-3. These results suggest that juniper oil may modulate the apoptosis through the activation of the interleukin-1-converting enzyme-like protease.