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신규향암제인 Heptaplatin의 인체 흑색종세포(SK-MEL-28)에 대한 세포생존률 및 유세포 분석
최수라,명평근 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8
Heptaplatin, cis-Malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolane]platinum(Ⅱ), is a novel platinum-based antitumor agent with clinical potential against human stomach cancer and the 3rd generation or the cisplatin. This study was performed to study how cisplain, heptaplatin and sunpla which is a mixture of heptaplatin and mannitol(w:w=1:2) affect cell viability of SK-MEL-28 human melanoma cell line. Heptaplatin(IC_50; 95.35㎛) and sun;la(IC_50;10.95㎛) were less effect than cisplatin (IC_50;10.92㎛) on the SK-MEL-28 cells. By cell cycle analysis using flow cytometry, it was identified that the cells were arrested at G2/M phase by cisplatin, heptaplatin and sunpla, and percentage of cell death group was increased according to increasing of time and concentration. These results suggest that cisplatin, heptaplatin and sunpla are a novel anticancer agent against human melanoma cell.
인체 흑색종 세포(SK-MEL-28 Cell Line)에서 Cisplatin, Heptaplatin, 그리고 Sunpla에 의한 Apoptosis의 유도
최수라,명평근 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8
A wide variety of cancer chemotherapeutic agents have been shown to induce programmed cell death (PCD, APOPTOSIS) in variou s tumor cell lines in vitro. cis-Malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isoprpopyl-1,3-dioxolane] platinum(Ⅱ) (heptaplatin), which is a new drug approved by KFDA in a novel platinum-based antitumor agent with clinical potential against stomach cancer and the 3rd generation of the cisplatin. This study was performed to know how heptaplatin and cisplatin and sunpla (mixture of heptaplatin and mannitol) affect on SK-MEL-28 cell line, and hour they induce the apoptosis. At EM analysis, the morphology of the cell was changed by treatment of the cisplatin, heptaplatin and sunpla. Apoptotic body formed around plasma membrane, and chromatin condensation represented in nucleus. This phenomenon is one of the characteristic of the apoptosis. The DNA of SK-MEL-28 cell line truncated by cisplatin and sunpla treatment was identified on 2% agarose gel electrophoresis. TUNEL assay was performed to know whether SK-MEL-28 cell die as apoptosis or necrosis by cisplatin, heptaplatin and sunpla. At this result, fluorescence intensity increased according to increase of time and concentration. Therefore, it was identified that cislatin, heptaplatin and sunpla induced apoptosis. Fas expressed on SK-MEL-28 cell membrane by cisplatin, heptaplatin and sunpla was identified by using flow cytometer and the expression of bcl-2(anti-apoptotic gene) decreased according to increase of concentration of the cisplation, heptaplatin and sunpla. Cisplatin, heptaplatin and sunpla induced apoptosis against 나-MEL-28 cell line, and the apoptotic mechanism was identified as Fas-mediated apoptosis and decreased bcl-2 expression.
토끼 귀의 출혈모델에서의 스트렙토키나제-덱스트란 포합체의 혈전용해효과 및 항원성 평가
명평근,최수라,박목순,박진규 충남대학교 약학대학 의약품개발연구소 2000 藥學論文集 Vol.16 No.-
The way in which a thrombolytic agent, streptokinase(SK, Dongkook) and streptokinase-Dextran conjugate(SK-DEX, Russia), cause the dissolution of haemostatic plugs of different ages was investigated in a novel rabbit bleeding model, and antigenicity of SK and SK-DEX was elvalutated by indirect ELISA using antisera collected after 7 days intravenous injection to a rabbit ear. Standardized incisions were made on rabbit ear and the wounds were left to heal for 0.5 h or 24 h, before the thrombolytic agent was infused. SK showed a pronounced fresh colt selectivity since it was significantly more effective in lysing flesh clots than old ones (96% vs 52%) respectively, percent lysis of haemostatic plus of different ages with SK-DEX conjugate (150,000 IU/㎏) was more effective in lysing fresh clots than old ones (20% vs 0%) respectively. The time onset of bleeding observed for SK(Dongkook) (19min) was the same as that of SK (Kabi) (19min), but the bleeding time observed for SK(Dongkook) (90 min) was twice longer than that formed SK(Kabi) (49min). However, the time to onset of bleeding of SK-DEX(Russia) (61min) was longer than that of the formed SK(Dongkook) (19 min). The bleeding time observed for SK-DEX(Russia, 150,000 IU/㎏) (1min) was shorter than that found for SK(Dongkook) (90min). The antigenicity of SK-DEX(Dongkook) was more effective than that of SK(Dongkook) in comparision with each control. These results suggest that the fresh clot selectivity demonstrated for SK and SK-DEX may be clinically beneficial and remains to be clarified by further invertigations.
Thiocholine ester 기질을 이용한 Acetylcholinesterase 활성부위의 구조특성
이천배,주은희,최수라,석대은,명평근 충남대학교 약학대학 의약품개발연구소 1999 藥學論文集 Vol.15 No.-
The inhibition pattern of three inhibitors(tacrine, decamethonium and propidium) on the hydrolysis of various thiocholine ester substrates by eel acetylcholinesterase was comparatively examined. When the substrate was acetylthiocholine, it showed a similar competitive inhibition by tacrine inhibitor, and a mixed type inhibition by decamethonium and propidium inhibitors. When the substrate was pentanoylthiocholine, it showed an uncompetitive inhibition by tacrine, and a noncompetitive inhibition by decamethonium. When the substrate was laurylthiocholine, it showed mixed type and uncompetitive inhibition by tacrine, and a competitive inhibition by decamethonium and propidium. Those results suggest that the active of acetylcholinesterase has the existence of hydorphobic site besides the anionic and esteratic site.
Thiocholine ester 기질을 이용한 Acetylcholinesterase 활성부위의 구조특성
이천배,주은희,최수라,석대은,명평근 충남대학교 기초과학연구소 1999 忠南科學硏究誌 Vol.26 No.1
The inhibition pattern of three inhibitors(tacrine, decamethonium and propidium) on the hydrolysis of various thiocholine ester substrates by eel acetylcholinesterase was comparatively examined. When the substrate was acetylthiocholine, it showed a similar competitive inhibition by tacrine inhibitor, and a mixed type inhibition by decamethonium and propidium inhibitors. When the substrate was pentanoylthiocholine, it showed an uncompetitive inhibition by tacrine, and a noncompetitive inhibition by decamethonium. When the substrate was laurylthiocholine, it showed mixed type and uncompetitive inhibition by tacrine, and a competitive inhibition by decamethonium and propidium. Those results suggest that the active site of acetylcholinesterase has the existence of hydrophobic site besides the anionic and esteratic site.
Choi, Su-La,Choi, Yun-Sil,Kim, Young-Kwan,Sung, Nack-Do,Kho, Chang-Won,Park, Byong-Chul,Kim, Eun-Mi,Lee, Jung-Hyung,Kim, Kyung-Mee,Kim, Min-Yung,Myung, Pyung-Keun 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10
We employed human SK-MEL-28 cells as a model system to identify cellular proteins that accompany N-(4-methyl)phenyl-O-(4-methoxy)phenyl-thionocarbamate (MMTC)-induced apoptosis based on a proteomic approach. Cell viability tests revealed that SK-MEL-28 skin cancer cells underwent more cell death than normal HaCaT cells in a dose-dependent manner after treatment with MMTC. Two-dimensional electrophoresis in conjunction with matrix-assisted laser desorption/ionization-time of flight (MALDl- TOF) mass spectrometry analysis or computer matching with a protein database further revealed that the MMTC-induced apoptosis is accompanied by increased levels of caspase-1, checkpoint suppressor-1, caspase-4, NF-κB inhibitor, AP-2, c-Jun-N-terminal kinase, melanoma inhibitor, granzyme K, G1/S specific eye/in D3, cystein rich protein, Ras-related protein Rab-37 or Ras-related protein Rab-13, and reduced levels of EMS (oncogene), ATP synthase, tyrosine-phosphatase, Cdc25c, 14-3-3 protein or specific structure of nuclear receptor. The migration suppressing effect of MMTC on SK-MEL-28 cell was tested. MMTC suppressed the metastasis of SK-MEL-8 cells. It was also identified that MMTC had little angiogenic effect because it did not suppress the proliferation of HUVEC cell line. These results suggest that MMTC is a novel chemotherapeutic and metastatic aoents aqainst the SK-MEL-28 human melanoma cell line.
Choi Su-La,Choi Yun-Sil,Kim Young-Kwan,Sung Nack-Do,Kho Chang-Won,Park Byong-Chul,Kim Eun-Mi,Lee Jung-Hyung,Kim Kyung-Mee,Kim Min-Yung,Myung Pyung-Keun The Pharmaceutical Society of Korea 2006 Archives of Pharmacal Research Vol.29 No.3
We employed human SK-MEL-28 cells as a model system to identify cellular proteins that accompany N-(4-methyl)phenyl-O-(4-methoxy)phenyl-thionocarbamate (MMTC)-induced apoptosis based on a proteomic approach. Cell viability tests revealed that SK-MEL-28 skin cancer cells underwent more cell death than normal HaCaT cells in a dose-dependent manner after treatment with MMTC. Two-dimensional electrophoresis in conjunction with matrixassisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis or computer matching with a protein database further revealed that the MMTC-induced apoptosis is accompanied by increased levels of caspase-1, checkpoint suppressor-1, caspase-4, NF-kB inhibitor, AP-2, c-Jun-N-terminal kinase, melanoma inhibitor, granzyme K, G1/S specific cyclin D3, cystein rich protein, Ras-related protein Rab-37 or Ras-related protein Rab-13, and reduced levels of EMS (oncogene), ATP synthase, tyrosine-phosphatase, Cdc25c, 14-3-3 protein or specific structure of nuclear receptor. The migration suppressing effect of MMTC on SK-MEL-28 cell was tested. MMTC suppressed the metastasis of SK-MEL-8 cells. It was also identified that MMTC had little angiogenic effect because it did not suppress the proliferation of HUVEC cell line. These results suggest that MMTC is a novel chemotherapeutic and metastatic agents against the SK-MEL-28 human melanoma cell line.