• 대한한의학방제학회지
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• 제26권3호
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• pp.195-205
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• 2018

Objectives : The purpose of this study was to observe the effects of Gardeniae Fructus on 4-HNE-induced apoptosis in PC-12 cell. Methods : A MTT assay was conducted to observe the cytotoxicity of Gardeniae Fructus on the PC-12 cell viability and the cytoprotective effects of Gardeniae Fructus on PC-12 cell against oxidative stress caused by 4-HNE. And western blot was conducted to observe the expression of Bax, Bcl-2, Caspase-3, $TNF-{\alpha}$ proteins which are involved in intrinsic and extrinsic apoptosis pathway. Results : 25, 50, 100, 200 and $400{\mu}g/m{\ell}$ of Gardeniae Fructus water extract had no cytotoxicity on PC-12 cell. $200{\mu}g/m{\ell}$ of Gardeniae Fructus water extract had significant cytoprotective effect on PC-12 cell against oxidative stress caused by 4-HNE. The expression of Bax protein in 50, 100 and $200{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly decreased in PC-12 cell. The expression of Bcl-2 protein in $200{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly increased in PC-12 cell. The expression of Caspase-3 protein in 100 and $200{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly decreased in PC-12 cell. The expression of $TNF-{\alpha}$ protein in $50{\mu}g/m{\ell}$ of Gardeniae Fructus was significantly decreased in PC-12 cell. Conclusions : These results suggest that Gardeniae Fructus water extract is effective to protect PC-12 cell from 4-HNE induced apoptosis.

• Journal of Acupuncture Research
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• 제22권6호
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• pp.37-50
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• 2005

Objectives : The purpose of this study was to investigate the anti-caner effect of Bee Venom and Melittin on the prostatic cancer cell(PC-3). The goal of study is to ascertain whether Bee Venom and Melittin inhibits the cell growth and cell cycle of PC-3, or the expression of relative genes and whether the regression of PC-3 cell growth is due to cell death or the expression of gene related to apoptosis. Methods : After the treatment of Pc-3 cells with Bee Venom and Melittin, we performed Fluorescence microscope, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify the cell viability, apoptosis and gene related to apoptosis. Results : 1. Compared with Control cell, the inhibition of cell growth reduced in proportion with the dose of Bee Venom or Melittin($0{\sim}10{\mu}g/ml$) in PC-3. 2. In PC-3, Cell viabilities of Bee Venom or Melittin treatment was decreased significantly. 3. The nucli of Control cells were stained round and homogenous in DAPI staining, but those of PC-3 were stained condense and splitted. 4. In PC-3, apoptosis of Bee Venom or Melittin treatment was increased significantly. 5. Bax, Caspase-3 and P ARP of Bee Venom or Melittin treatment was increased significantly and Bcl-2 of Bee Venom or Melittin treatment was decreased significantly. Caspase-9 of Bee venom treatment was increased significantly. Conclusion : These results indicate that Bee Venom and Melittin inhibits the growth of prostate cancer cells, has anti-cancer effects by inducing apoptosis. We wish that the anti-cancer effects of Bee Venom and Melittin are used to clinical caner treatment.

• 약학회지
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• 제43권3호
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• pp.369-377
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• 1999

A new series of highly water soluble platinum(II) complexes {Pt(II)[1,3-bis(phenylthio) propane](trans- -1,2-diaminocyclohexane) (PC-1) and Pt(II)[1,3-bis-(phenythio)propane] cis-1,2-diaminocyclohexane(PC-2)} were synthesized, and characterized by their elemental analysis and by various spectroscopic techniques[infrared(IR), 13C-nuclear magnetic resonance (NMR)]. In vitro antitumor activity of new Pt(II) complexes was tested against P-388 and L-1210 mouse lymphocytic leukemia cell lines, PC-14 / P, PC-14/ADM and PC-14 / CDDP human pulmonary adenocarcinima, DU-145 human prostate carcinoma, HT-1376 human bladder carcinoma, ZR-75-1 human breast carcinoma, MKN-45/P and MKN-45/CDDP human gastric adenocarcinoma cell lines using colorimetric MTT[3-(4,5-dimethyl thiazol-2-yl)-2.5-diphenyltetrazoliumbromide] assay for cell survival and proliferation. PC-1 showed active against L-1210, P-388 leukemia, human lung, stomach, prostate, bladder and breast cancer cell lines, and the antitumor activity of these compounds were comparable or superior to those of PC-2 and displatin. The nephrotoxicities of PC-1 and PC-2 were found quite less than that of cisplatin using MTT and [3H] thymidine uptake in rabbit proximal tubule cells and human kidney cortical cells. Based on these results, this novel platinum (II) complex compound (PC-1) represents a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low nephrotoxicity.

• KSBB Journal
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• 제18권5호
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• pp.424-428
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• 2003

내분비계장애물질인 bisphenol A, nonylphenol, pentachlorophenol을 대상으로 여성 유방암세포 유래 MCF-7 세포주와 남성 전립선암세포 유래 PC-3 세포주에서 세포 증식효과를 MTT 방법으로 조사하였다. MCF-7 세포주에 이들 세 종류의 내분비계장애물질을 농도별로 처리하여 세포증식에 미치는 영향을 조사한 결과 모두 세포증식을 촉진하는 결과를 보였다. $10^{-7}$ M에서 $10^{-6}$ M 농도 범위에서 MCF-7 세포의 최대증식효과를 유도하였다. 그러나 PC-3 세포주의 경우에는 세포증식에 bisphenol A, nonylphenol, pentachlorophenol 모두 영향을 미치지 못하였다. 이러한 결과는 이들 세 종류의 내분비계 장애물질이 남성 전립선세포 유래인 PC-3 세포주의 증식에는 관여하지 않고 여성 유방암 세포에서 유래하고 에스트로젠 반응성인 MCF-7 세포주에만 증식효과를 갖는 사실을 보여주고 있으며 이는 bisphenol A, nonylphenol, pentachlorophenol이 여성호르몬인 에스트로젠과 유사한 역할을 한다는 사실을 보여주는 것이라 할 수 있다.

• 생명과학회지
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• 제21권11호
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• pp.1573-1578
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• 2011

Silibinin은 milk thistle에서 분리된 주된 생리활성 성분으로 강력한 항산화제, 항암활성에 대해서 보고되어 있다. 하지만 항암활성에 대한 정확한 기전에 대해서는 밝혀져 있지 않다. 본 연구에서는 인간 전립선 암세포주인 PC-3 세포를 이용하여 세포사멸의 기전을 조사하였다. MTT assay를 통해 세포독성을 확인하였고, PI 염색을 통해 세포주기를 확인하고, Annexin-V/PI 염색을 통한 세포사멸을 확인하였다. 뿐만 아니라 western blot을 이용하여 세포주기 및 세포사멸에 관련된 단백질 발현 정도를 확인하였다. 본 연구의 결과에서 silibinin은 인간 전립선 암세포주인 PC-3 세포에서 세포주기관련 단백질의 발현을 조절하여 세포주기 진행을 억제함으로써 세포사멸을 유도 함을 알 수 있었다.

• Journal of Animal Science and Technology
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• 제48권5호
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• pp.719-726
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• 2006

본 실험은 가수분해 시킨 유청단백질과 Lb. casei HY2782의 세포 추출물을 인간 전립선 세포에처리하여 세포 내 glutathione 농도 상승효과와 산화제 tbutyl hydroperoxide (TBHP)에 의한핵산 손상 방지 효과를 알아보기 위한 실험으로 전립선 세포 RWPE1 cells와 PC3MMM2 cells에 hydrolyzed WPI(500g/m)를 48시간 동안 처리 시, 가수분해 시키지 않은 WPI 보다 각각 28.2%, 38.4%씩 GSH 농도가 증가하는 것을 유의적으로 확인할 수 있었고(p<0.05), 카제인의 경우 RWPE1 cells and PC3MMM2 cells에 처리 시 가수분해 시킨 카제인과의 유의차가 보이지 않았으며 (p<0.05), glutathione 합성저해제인 Buthionine sulfoximin (BSO) 처리 시 Glutathione 농도가 현저하게 낮아지는 것을 확인할 수가 있었다(Anderson, 1998). 전립선 세포 RWPE1 cells 와 PC3MMM2 cells에 Lb. casei HY2782 세포 추출물 처리 시 PC3MMM2 cell에서는 GSH의 농도가 유의적으로 증가하는 것을 확인 할 수 있었고(p<0.05), RWPE1 cell에서는 증가 성향을 확인 할 수가 있었다.가수분해 시킨 유청단백질과 Lb. casei cell 세포추출물 처리 시 산화제에 의한 세포의 DNA 손상 억제정도와 사멸율의 변화를 확인 한 결과, 가수분해 WPI를 처리 후 Oxidant tbutyl hydroperoxide (TBHP)(500mM)를 이용하여 PC3MMM2 세포에 산화적인 스트레스를 주었을 시, TBHP만 처리한 구에서는 62.0%의 생존율을 나타내었고, glutathione 합성저해제인 BSO를 첨가시킨 처리구에서는 33.7%의 생존율을 나타내었다. 가수분해 시킨 WPI와 카제인 처리구에서는 각각 86.7%, 63.6%의 생존율을 나타내었고, WPI와 BSO를 함께 처리한 구에서는 71.5%의 생존율을 나타내었다. 이는 BSO에 의해 GSH의 생성이 저해되어 나타난 결과로 보여지며(Anderson, 1988), 가수분해시킨 WPI 가 가수분해 시킨 카제인보다는 높은 생존율을 나타내는 것을 유의적으로 확인할 수가 있었다. (p<0.05) Lb. casei 세포 추출물처리에 의한 생존율에서는 가수분해시킨 WPI와 유사한 수준의 82.4%의 생존율을 보였다. 가수분해시킨 유청단백질과 Lb. casei cell 세포추출물에 의한 세포에서의 glutathione 농도 증가에 의해 산화제에 의한 DNA의 손상이 적었던 것으로 사료되어지고, 이로 인해 생존율이 높았던 것으로 판단된다.

• 대한본초학회지
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• 제30권1호
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• pp.59-65
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• 2015

Objectives : The purpose of this study was to observe the effects of Polygalae Radix(PR) on 4-HNE-induced apoptosis in PC-12 cell. Methods : A MTT assay was conducted to observe the cytotoxicity of Polygalae Radix on the cell viability and the cytoprotective effect of Polygalae Radix against 4-HNE that causes oxidative stress-induced cytotoxicity, and then a western blot was conducted to observe the expression of $TNF-{\alpha}$, caspase-3, Bax and Bcl-2 protein that are important factors involved with apoptosis signaling pathway. Results : The Polygalae Radix water extract $25{\mu}g$, $50{\mu}g$, $100{\mu}g$ and $200{\mu}g/mL$ had no cytotoxicity on the PC-12 cell. The Polygalae Radix water extract $25{\mu}g$, $50{\mu}g$ and $100{\mu}g/mL$ had the cytoprotective effect against 4-HNE that causes cytotoxicity on the PC-12 cell. The Polygalae Radix water extract $50{\mu}g/mL$ significantly suppressed the increase in $TNF-{\alpha}$ protein expression in PC-12 cell. The Polygalae Radix water extract $25{\mu}g$ and $50{\mu}g/mL$ significantly suppressed the increase in caspase-3 protein expression in PC-12 cell. The Polygalae Radix water extract $25{\mu}g$, $50{\mu}g$ and $100{\mu}g/mL$ suppressed the increase in Bax protein expression in PC-12 cell but had no significance. The Polygalae Radix water extract $25{\mu}g$ and $100{\mu}g/mL$ significantly prevented the decrease in Bcl-2 protein expression in PC-12 cell, Conclusions : These results suggest that the Polygalae Radix water extract is effective in inhibiting apoptosis.

• 생물정신의학
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• 제5권2호
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• pp.219-226
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• 1998

Cytosine arabinoside(AraC) inhibits DNA synthesis and ${\beta}$-DNA polymerase, an enzyme involved in DNA repair. This, a potent antimitotic agent, is clinically used as an anticancer drug with side effect of severe neurotoxicity. Earlier reports suggested that inhibition of neuronal survival by AraC in sympathetic neuron may be due to the inhibition of a 2'-deoxycytidine-dependent process that is independent of DNA synthesis or repair and AraC induced a signal that is triggers a cascade of new mRNA and protein synthesis, leading to apoptotic cell death in cultured cerebellar granule cells. The present study would suggest whether caspase family(ICE/CED-3-like protease) involved in AraC-induced apoptosis pathway of PC12 cells. It was observed that treatment of PC12 cells with AraC led to decrease of viability by MTT assay and morphology changes, which did not suggest that AraC induced apoptosis in PC12 cells. The mRNA of caspase-1/caspase-3 were expressed in PC12 cells constitutively, and AraC did not activate caspase family. These results suggest that caspase-1/caspase-3 may not be required for AraC-induced cell death pathway in PC12 cells.

• Biomolecules & Therapeutics
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• 제12권1호
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• pp.9-18
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• 2004

Opening of the mitochondrial permeability transition pore has been recognized to be involved in cell death. The present study investigated the effect of ${\beta}$-carbolines (harmaline and harmalol) and deprenyl on the dopamine-induced change in the mitochondrial membrane permeability and cell death in differentiated PC12 cells. Cell death due to 250 4{\mu}$M dopamine was inhibited by caspase inhibitors (z-IETD.fmk, z-LEHD.fmk and z-DQMD.fmk) and antioxidants (N-acetylcysteine, ascorbate, superoxide dismutase, catalase and carboxy-PTIO). ${\beta}$-Carbolines prevented the dopamine-induced cell death in PCl2 cells, while deprenyl did not inhibit cell death. ${\beta}$-Carbolines decreased the condensation and fragmentation of nuclei caused by dopamine in PC12 cells. ${\beta}$-Carbolines inhibited the decrease in mitochondrial transmembrane potential, cytochrome c release, formation of reactive oxygen species and depletion of GSH caused by dopamine in PC12 cells, whereas deprenyl did not decrease dopamine-induced mitochondrial damage. ${\beta}$-Carbolines, deprenyl and antioxidants depressed the formation of nitric oxide and melanin in dopamine-treated PC12 cells. The results suggest that cell death due to dopamine PC12 cells is mediated by caspase-8, -9 and -3. Unlike deprenyl, ${\beta}$-carbolines may attenuate the dopamineinduced cell death in PC12 cells by suppressing change in the mitochondrial membrane permeability through inhibition of the toxic action of reactive oxygen and nitrogen species.

• Biomolecules & Therapeutics
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• 제11권2호
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• pp.91-98
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• 2003

A new series of highly water soluble platinum(II) complexes[Pt(II)(DL-2-hydroxy-3-methylbutyrate)(trans-l-1,2-dimninocyc1ohexane)] (PC-1) and [Pt(II)DL-2-hydroxy-3-methylbutyrate](cis-1,2-diaminocyclohexane)](PC-2) were synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared(IR), $^{13}C$-nuclear magnetic resonance (NMR)]. In vitro antitumor activity of new Pt(II)complexes was tested against MKN-45, MKN/ADM and MKN/CDDP human gastric adenocarcinoma cell lines using colorimetric MTT[3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyltetrazoliumbromide] assay for cell survival and proliferation. PC-1 and PC-2 showed active against MKN-45/P, MKN/ADM and MKN/CDDP human gastric cancer cell lines, and the antitumor activity of these compounds were comparable or superior to that of cisplatin. The nephrotoxicities of PC-1 and PC-2 were found quite less then that of cisplatin using MTT and [$^3H$] thymidine uptake tests in rabbit proximal tubule cells, human kidney cortical cells human renal cortical tissues. Based on these results, these novel platinum(II) complex compounds(PC-1 ＆ PC-2) represent a valuable lead in the development of the new anticancer chemotherapeutic agents capable of improving antitumor activity and low nephrotoxicity.