• 동의생리병리학회지
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• 제17권2호
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• pp.293-296
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• 2003

Prunellae Spica is a flower petal of Prunella vulgaris var. lilacina used for treatment of lymphoma, breast cancer, hepatitis and pathological fluid related diseases in oriental medicine. We tried to evaluate the mechanism of Prunellae Spica in the treatment of cancer. The ethyl acetate layer of Prunellae Spica showed a good cytotoxicity on U937 cells with IC50 of 8 ug/ml. It induced apoptosis in U937 dose-dependently by cell cycle analysis following PI staining. We also confirmed it induced DNA fragmentation in U937 cells from the concentration of 10 ug/ml. From western blot assay we observed the ethyl acetate layer of Prunellae Spica downregulated procaspase-3 and cleaved PARP in a dose dependent manner, whereas it didn't affect bax and bcl-2. Taken together, these results indicate the ethyl acetate layer of Prunellae Spica can induce apoptosis in U937 cells suggesting it can be potently applied to cancer.

• 동의생리병리학회지
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• 제19권3호
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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.

• 대한본초학회지
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• 제33권4호
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• pp.69-76
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• 2018

Objectives : Dracocephalum palmatum Stephan (Lamiaceae) is a medicinal plant used by the East-Russian nomads but there were few studies on this plant. This study was to evaluate anti-cancer effects of D. palmatum Stephan leaf hexane fraction on human derived prostate cancer cell death. Methods : The dried leaves of D. palmatum were dissolved in methanol, and hexane fraction (DpLH) was again obtained from lyophilized methanol extract (DpLM). DpLH was investigated by measuring by MTT assay and annexin V/PI staining to evaluate its effects on the cell viability and apoptosis of PC-3 cells. The ROS generations were detected by DCF-DA dye. The protein expressions were confirmed by p-AKT, Bcl-2, Bax, procaspase-3 activities. Results : After treatment of DpLH to PC-3 cells, the cell proliferation was significantly inhibited, and in addition, DpLH treatment also accelerated apoptosis of PC-3 cells. When DpLH was treated to the PC-3 cells, its ROS production significantly decreased. The proportion of all proteins (p-AKT/actin, Bcl-2/Bax and procaspase-3/actin ratios) showed decreasing tendency of expression compared with the control group. Conclusions : As shown in the above results, the extract from D. palmatum inhibits ROS production and promotes cell death, which is considered to be a relatively safe induction of cell death when administered to a living body. In conclusion, these results suggested that DpLH may have anti-cancer effect in human prostate cancer cell.

• 대한한방내과학회지
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• 제35권1호
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• pp.79-91
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• 2014

Objectives : To investigate the anti-cancer effect of Palbohoichoon-tang (PBHCT) extracts. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were microscopically analyzed after staining with $10{\mu}M$ 2-[4-amidinophenyl]-6-indolecarbamidine dihydrochloride (DAPI) and TUNEL. We also analyzed expression of Bcl2, $Bcl_{xL}$, Bax, procaspase-3, procaspase-9, and procyclic acidic repetitive protein (PARP) by western blot method. Results : Observations showed that PBHCT induced the apoptotic cell death proved by increased sub-G1 phase cell population, apoptotic body formation and chromatin condensation. Western blot analysis of total cell lysates revealed that the PBHCT induced cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP). In addition, PBHCT dose-dependently increased the activity of caspase-9, caspase-3 and PARP-1. Furthermore, PBHCT reduced anti-apoptotic Bcl2, $Bcl_{xL}$ expression which contributed to the loss of mitochondrial membrane potential and the activations of caspase-9 and caspase-3. Conclusions : These findings suggest that PBHCT exerts anti-cancer effects on human neuroblastoma SH-SY5Y cells by inducing apoptotic death via down-regulation of anti-apoptotic proteins such as Bcl2 and $Bcl_{xL}$, up-regulation of pro-apoptotic proteins such as Bax, and activation of caspase cascades and PARP-1.

• 생약학회지
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• 제44권3호
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• pp.269-274
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• 2013

In this study, we investigated the anticancer effect of rice bran extract in HL-60 human promyelocytic leukemia cells. The extract of rice bran inhibited the proliferation of HL-60 cells. When treated with the rice bran extract, we could observe the apoptotic characteristics such as apoptotic bodies and the increase of sub-G1 hypodiploid cell population, increase of Bax level, decrease of Bcl-2 expression, cleavage of procaspase-3, cleavage of procaspase-9 and cleavage of poly(ADP-ribose) polymerase(PARP) in HL-60 cells. Furthermore, the apoptosis induction of HL-60 cells treated with the rice bran extract was also accompanied by the inactivation of mitogen-activated protein kinases (MAPK) such as ERK1/2 MAPK and p38 MAPK. In addition, the rice bran extract induced the down-regulation of c-myc. These data suggested that the rice bran extract could induce the apoptosis via the inactivation of ERK1/2 MAPK and p38 MAPK, and the down-regulation of c-myc in HL-60 acute pomyelocytic leukemia cells. The results support that the rice bran extract might have potential for the treatment of acute promyelocytic leukemia.

• Toxicological Research
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• 제18권3호
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• pp.275-283
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• 2002

Fungal metabolite, gliotoxin is an epipolythiodioxopiperazin (ETP) class and has various roles including immunomodulatory and apoptotic effects. This study was designed to evaluate the mechanism by which gliotoxin exerts the apoptosis on human promyelocytic leukemic HL-60 cells. Herein, we demonstrated that the gliotoxin decreased the cell viability in a time-dependent manner Gliotoxin-induced cell death was confirmed us apoptosis characterized by chromatin condensation and ladder-pattern fragmentation of genomic DNA. Gliotoxin increased the catalytic activities of caspase-3 and caspase-9. Activation of caspase-3 was further confirmed by degradation of procaspase-3 and poly(ADP-ribose) polymerase (PARP) by gliotoxin in HL-60 cells. Furthermore, gliotoxin induced the changes of mitochondrial transmembrane potential (MTP). Antioxidants, including GSH and NAC, markedly inhibited apoptosis with conistent suppression of enzymatic activity of caspase-3, caspase-9, and MTP loss in gliotoxin-treated cells. Taken together, we suggest that gliotoxin function as an oxidant and ploys proapoptotic roles in HL-60 cells via activation of intrinsic caspase cascades as well as mitochondrial dysfunction.

• 생명과학회지
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• 제26권4호
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• pp.398-405
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• 2016

3-Hydroxy-3-methylglutaryl coenzyme A 환원효소의 억제제로 알려진 statin은 고지혈증 치료제로 널리 사용되고 있고, 또한 다양한 암에서 항암효과를 나타낸다고 알려져 있다. 최근 연구에서는 reactive oxygen species (ROS)가 세포사멸 신호에 중요한 역할을 한다고 보고하였지만, rosuvastatin에 의한 ROS 생성의 정확한 기전은 아직 밝혀지지 않았다. 인간 전립선암 세포주인 PC-3 세포를 이용하여 rosuvastatin에 의한 세포사멸 경로를 확인하였다. 세포독성, 세포사멸과 ROS의 생성을 측정하기 위해서 MTT assay, annexin V/PI 염색과 DCFH-DA염색을 통해 flow cytometry에 의해 측정하였다. 본 연구의 결과에서, rosuvastatin은 농도와 시간 의존적으로 세포 생존율 감소와 세포형태변화를 확인할 수 있었다. Flow cytometry 분석을 통해 세포주기와 apoptosis를 확인하였고 Western blotting assay를 통하여 PARP와 procaspase-3가 감소되는 것을 통해 apoptosis를 재확인 할 수 있었다. 또한 rosuvastatin은 농도 의존적으로 ROS 생산을 증가하였고, ROS 생성 저해제인 N-acetylcysteine (NAC) 처리를 통해 ROS와 apoptosis가 회복되었다. 따라서 rosuvastatin이 ROS 생성을 통해 apoptosis를 유도한다는 것을 알 수 있었고, 이는 인간 전립선 암세포에 대한 항암치료제로서의 가능성을 제시한다.

• Toxicological Research
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• 제19권3호
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• pp.197-203
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• 2003

Apicidin, a histone-deacetylase inhibitor, has been successfully used to inhibit the growth of cancer cells. In this study, the apoptotic potential and mechanistic insights of apicidin were investigated in human myeloid leukemia U937 cells. Treatment of U937 cells with apicidin resulted in a decrease of cell viability with apoptotic characteristics, including chromatin condensation and ladder-pattern fragmentation of genomic DNA. Apicidin converted the procaspase-3 protease to catalytically active effector protease, resulting in subsequent cleavage of poly (ADP-ribose) polymerase (PARP) and inhibitor of caspase-activated deoxyribonuclease (ICAD). In addition, apicidin induced the activation of caspase-9 protease and the cytosolic release of mitochondrial cytochrome c with mitochon-drial membrane potential transition. Moreover, apicidin transiently increased the expression of Fas and Fas ligand proteins. Taken together, the results suggest that apicidin induces apoptosis of U937 cells through activation of intrinsic caspase cascades and Fas/FasL system with mitochondrial dysfunction.

• 생명과학회지
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• 제27권2호
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• pp.164-171
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• 2017

본 연구에서는 인간 자궁경부암세포주에서 S-allylcysteine (SAC)이 세포자멸경로에 중요한 역할을 담당하는 initiator caspase의 하나인 caspase-9와 effector caspase에 속하는 caspase-3 및 caspase-7 그리고 DNA 복구에 관여하는 poly ADP-ribose polymerase (PARP)의 발현조절에 미치는 영향과, SAC에 의한 이러한 세포자멸 및 DNA 복구 관련 단백질의 발현변화가 세포증식억제를 통한 기능적 작용을 유발하는지를 조사하였다. 단백질 발현분석 결과, 특히 50 mM의 SAC로 48시간 동안 처리하였을 경우, procaspase-3, -7, -9 및 PARP의 발현은 각각 94%, 38%, 95% 및 64% 감소되었으며, 이와 반대로 caspase-3, -7, -9 및 cleaved-PARP의 발현은 현저히 증가되었다. 또한 cell proliferation assay 결과, 20 mM 이상의 SAC 처리는 6, 12, 24 및 48시간에서 농도 및 시간 의존적인 세포증식 억제효과를 나타내었다. 이러한 결과는 SAC 처리가 자궁경부암세포의 증식을 억제하며, 이에 대한 가능한 분자적 작용기전들 중의 하나로 세포자멸과정 중 initiator caspase의 하나인 caspase-9의 활성을 유도하고 이에 따른 effector caspase인 caspase-3과 caspase-7의 활성을 촉진시킬 뿐만 아니라 DNA 복구에 관여하는 PARP의 불활성화를 초래함으로써 세포자멸 유도에 관여하는 것으로 사료된다.

• 생명과학회지
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• 제27권3호
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• pp.267-274
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• 2017

본 연구는 말기 신부전 환자의 체내에서 증가되는 시안산이 신경학적 합병증의 원인으로 작용하는지 알아보고자 시안산 처리에 따른 신경아교종 세포인 C6 세포의 변화를 관찰하였다. 또한, 시안산에 의해 발현되는 세포자멸사 관련 인자를 알아보기 위하여 western blot 및 유전자 발현의 변화를 검색하기 위하여 cDNA 유전자 미세배열분석을 하였다. 시안산의 처리 농도가 0, 1, 5, 10, 20, 40 mM 증가할수록 신경아교종 세포의 생존율이 유의하게 감소하였고 세포자멸사에 주된 역할을 하는 caspase-8는 증가되었고 procaspase-3는 감소하였다. 그러나 caspase-8에 의해 활성화되는 Bax 단백질은 시안산의 처리 농도가 증가할수록 caspase-8의 증가에도 감소하였고, 세포자멸사를 조절하는 단백질인 Bcl-2와 IAP은 명확히 확인할 수 없었다. cDNA 유전자 미세배열 분석 결과, 총 1,099 종의 유전자 중에서 934 개의 유전자가 감소하였고 증가된 것은 165 개였다. 세포자멸사 관련 유전자에서도 감소한 것은 16 개였고, 증가된 6 개 유전자 가운데 heat shock 70 kD protein 1A가 현저한 증가를 나타내었다. 이상의 결과로 보아, 시안산은 신경아교종 세포에서 caspase-8 및 caspase-3와 관련된 세포자멸사를 유발시키며, 신경아교종 세포의 유전자들의 발현을 감소시키는 것으로 생각된다. 따라서 체내에서 증가된 시안산이 신경아교종 세포에 영향을 미쳐 말기 신부전 환자의 뇌병증에도 영향을 주는 것이라 생각된다.