• 제목/요약/키워드: SK-HEP-1 cells

검색결과 55건 처리시간 0.027초

천연 식물 추출물에서 Matrix Metalloproteinase-9 활성 억제제의 분리 및 특성화에 관한 연구 (A study on separation and characterization of matrix metalloproteinase-9 inhibitors from natural plants)

  • 허용철;박성우;김태진
    • 분석과학
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    • 제18권3호
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    • pp.188-193
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    • 2005
  • 천연 식물 추출물로 처리한 matrix metalloproteinase (MMP)의 저해능 실험에서 귀전우 (Euonymus alatus)의 methanol 가용성 분획이 matrix metalloproteinase (MMP-9) 활성에 저해능이 있는 것으로 나타났다. Silica gel column에서 ethyl acetate와 hexane의 혼합비를 달리하는 방법으로 다섯 개의 부분으로 분획하였고 그 중에서 세 개의 분획이 MMP-9 활성에 저해능이 있는 것으로 나타났다. 인체 간암 세포인 Hep3B 세포주와 인체 정상 세포인 Chang 세포주를 같은 조건으로 키운 후 형태학적 변화를 관찰한 결과, 귀전우의 메탄올 추출물로 처리한 세포에서만 핵의 주위에 검은 반점이 넓게 분포되어 있는 것이 관찰되었다. 인체 정상 세포주에 대한 독성 확인 실험에서 대부분의 메탄올 가용성 분획은 독성을 나타내지 않는 것으로 나타났다.

Production of Lignin Peroxidase by Phellinus igniarius and Cytotoxic Effects of Lignin Hydrolysates Derived from Wood Biomass on Cancer Cells

  • Lee, Jae-Sung;Lee, Jong-Suk;Yoon, Jae-Don;Beak, Sung-Mok;Bosire, Kefa-O.;Lee, Yong-Soo;Kim, Jung-Ae
    • Biomolecules & Therapeutics
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    • 제12권3호
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    • pp.189-193
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    • 2004
  • Over the past several years, research efforts have been directed both at economically producing valuable substances from the wood biomass and at producing lignolytic enzymes at a lower cost. In the present study, we found that Phellinus igniarius, the basidiomycetes, secreted lignin peroxidase as a main lignolytic enzyme, which was detected maximum activity at 16th day of culture and showed 37 kDa of molecular mass in identification by activity assay and purification by anion-exchange chromatography. The Phellinus igniarius-derived lignin peroxidase hydrolyzed steam-exploded wood (Quercus mongolica) powder into small molecules showing cytotoxicity against cancer cel1s (HepG2 hepatoma, SK-N-SH neuroblastoma, B16 melanoma, MBT-2 bladder cancer). In addition, the enzyme hydrlysates of lignins (ELg) that were extracted from the steam-exploded oak showed more potent cytotoxic effects on the cancer cells than the enzyme hydrolysates of wood biomass (EWp), indicating that the cytotoxic effect of EWp may be due to the enzyme-degraded products of lignin among the lignocellulosics. Furthermore, the cytotoxic effect of ELg on Chang, normal liver cells, was much less potent than that of ELg on HepG2 and B16 cancer cells, indicating that the cytotoxic effect of ELg may be specific for cancer cells. The present results suggest that Phellinus igniarius may be a useful resource for the large-scale production of lignin peroxidase and that the lignin peroxidase may be applied for the generation of valuable biodegradation products from wood lignocellulosics for medical use.

정상 간세포와 간암세포의 플라즈마 특성에 관한 비교연구 (Comparative study of plasma effects on human liver normal and cancer cells)

  • 김대연;권보미;김단비;최원호;신현정
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2008년도 추계학술대회A
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    • pp.1539-1542
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    • 2008
  • Plasma is 4th state of matters, which consists of electrons, neutral, and ionized particles. In biomedical research, cold plasma, which is generated in atmospheric condition, has been applied to disinfect microorganisms such as bacteria and yeast cells. Because of its low temperature condition, the heat-sensitive medical device can be easily sterilized by the cold plasma treatment. In recent years, the effects of plasma on mammalian cells have arisen as a new issue. Generally, plasma induces intensity dependent necrotic cell death. In this research, we investigate the feasibility of cold plasma treatment for cancer therapy by conducting comparative study of plasma effects on normal and cancer cells. We use THLE-2 (human liver normal cell) and SK-Hep1 (human liver metathetic cancer cell) as our target cells. The needle type of cold plasma is generated by the Helium plasma device. Two types of cells have different onset plasma conditions for the necrosis, which may be explained by difference in electrical properties of these two cell types.

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Caspase-3 Specifically Cleaves $p21^{WAF1/CIP1}$ in the Earlier Stage of Apoptosis in SK-HEP-1 Human Hepatoma Cells

  • Park, Jeong-Ae;Kim, Kyu-Won;Kim, Shin-Il;Lee, Seung-Ki
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1998년도 Advances in Ginseng Research - Proceedings of the 7th International Symposium on Ginseng -
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    • pp.231-243
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    • 1998
  • In the present study, we provide evidence that ginsenoside $Rh_2$ (G-$Rh_2$) as well as staurosporine induces apoptosis of human hepatoma SK-HEP-1 cells by caspase 3-mediated processing of $p21^{WAFI/CIPI}$ in the early stage of apoptosls. Immunoblottings showed that G-$Rh_2$ as well as statrosporine induced the processing of caspase-3 to an active form, pl7. In stable Bcl-2 transfectants however, G-$Rh_2$ induced DNA fragmentation, while staurosporine did not. In the early stage of apoptosis, $p21^{WAFI/CIPI}$ was detected to undergo proteolytic processing specifically conducted by caspase-3. $p21^{WAFI/CIPI}$ translated in vitro was cleaved into a p14 fragment, when incubated with cell extracts obtained from either G-$Rh_2$- or staurosporine-treated cells. Cleavage was equally inhibited in both cases by adding Ac-DEVD-cho, a specific caspase-3 inhibitor, but not by Ac-YVkD-cho, a specific caspase-l inhibitor. Similarly, $p21^{WAFI/CIPI}$ was efficiently leaved by recombinant caspase-3 overexpressed in E. coli. Moreover, the endogenous $p21^{WAFI/CIPI}$ of untreated-cell extracts was also cleaved by recombinant caspase-3. Mutation analysis allowed identification of two caspase-3 cleavage sites, $DHVD^{112}$/L and $SMTD^{149}$/F, which are located within, or near the interaction domains for cyclins, Cdks, and PCNA. Taken together, these results show that G-$Rh_2$ as well as staurosporine increases caspase-3 activity, which in turn directly cleaves $p21^{WAFI/CIPI}$ resulting in elevation of Cdk kinase activity in the early stages of apoptosis. We propose that proteolytic cleavage of $p21^{WAFI/CIPI}$ is a functionally relevant event that allows unleashing the cyclin/Cdk activity from the inhibitor seen in the earlier stage of apoptosis, the event of which may be associated with the triggering mechanism for the execution of apoptosis.

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Protective effects of synthetic of 3-Alkoxy-6-allylthiopyridazine against aflatoxin $B_1$- induced hepatotoxicity

  • Soon, Shin-Hea;Kang, Joo-Yeon;Park, Myung-Sook;Kwon, Soon-Kyoung
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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    • pp.172.2-173
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    • 2003
  • 3-Alkoxy-6-allylthiopyridazine derivatives showed the strongest protective effect against oxidative stress and their anticancer effect determined on the growth of SK-Hep-l hepatocellular carcinomar cells. The allythio group as a pharmacologically active group was introduced into pyridazine nucleus and a substituent such as halogen or alkoxy was also introduced into paraposition of allylthio group. Five kinds of 3-alkoxy-6-allylthiopyridazine derivatives were synthesized and their chemoprotective activities examined in rats exposed to aflatoxin $B_1$-toxicant. (omitted)

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Dimethyl sulfoxide elevates hydrogen peroxide-mediated cell death in Saccharomyces cerevisiae by inhibiting the antioxidant function of methionine sulfoxide reductase A

  • Kwak, Geun-Hee;Choi, Seung-Hee;Kim, Hwa-Young
    • BMB Reports
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    • 제43권9호
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    • pp.622-628
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    • 2010
  • Dimethyl sulfoxide (DMSO) can be reduced to dimethyl sulfide by MsrA, which stereospecifically catalyzes the reduction of methionine-S-sulfoxide to methionine. Our previous study showed that DMSO can competitively inhibit methionine sulfoxide reduction ability of yeast and mammalian MsrA in both in vitro and in vivo, and also act as a non-competitive inhibitor for mammalian MsrB2, specific for the reduction of methionine-R-sulfoxide, with lower inhibition effects. The present study investigated the effects of DMSO on the physiological antioxidant functions of methionine sulfoxide reductases. DMSO elevated hydrogen peroxide-mediated Saccharomyces cerevisiae cell death, whereas it protected human SK-Hep1 cells against oxidative stress. DMSO reduced the protein-carbonyl content in yeast cells in normal conditions, but markedly increased protein-carbonyl accumulation under oxidative stress. Using Msr deletion mutant yeast cells, we demonstrated the DMSO's selective inhibition of the antioxidant function of MsrA in S. cerevisiae, resulting in an increase in oxidative stress-induced cytotoxicity.

칠황버섯으로부터 분리한 Lanostane-type Triterpenoid의 암세포성장 억제효과 (Cancer Cell Growth Inhibition of Lanostane-type Triterpenoids Isolated from Ganoderma gibbosum)

  • 김동화;이상국;박희준
    • 생약학회지
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    • 제51권1호
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    • pp.36-40
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    • 2020
  • The CHCl3 fraction of the MeOH extract of Ganoderma gibbosum (Ganodermataceae) exhibited cytotoxic activity on five cancer cell lines (MDA-MB-231, SK-hep1, A549, HCT116, and SNU638). Six highly oxygenated lanostane-type triterpenoids (lanostanoids) were isolated by column chromatography to test cytotoxicity on cancer cells. The five known lanostanoids were identified as gibbosic acids A, B, D, G, and H by comparison of molecular ion peaks with the literature data. The structure of a new lanostanoid, gibbosic acids I, was identified to be 3β,8β,15β,20S-tetrahydroxy-7,12,23-trioxolanost-9(11)-en-26-oic acid on the basis of NMR and MS spectroscopy. The three lanostanoids of gibbosic acids A, H, and I of the six isolates significantly suppressed the growth of cancer cells. In particular, the IC50 of gibbosic acid H was prominently low ranging from 2.64-6.56 μM.

Conditioned Media of ASC-17D Sertoli Cells Induce G1-Growth Arrest of DU145 Human Prostate Cancer Cells

  • Kang, Hyeog;Kang, Sang-Wook;Choi, Sang-Hyun;Lee, Kwang-Ho;Chun, Boe-Gwun;Min, Bon-Hong
    • BMB Reports
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    • 제31권5호
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    • pp.508-514
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    • 1998
  • We studied the effects of ASC-17D rat Sertoli cell-conditioned media (rSCCM) on the proliferation of the DU145 prostate cancer cells. rSCCM was prepared from ASC-17D cells cultured in DMEM/F-12 serum-free media at a nonpermissive temperature of $40^{\circ}C$, which is the condition for the high expression of c1usterin. We found that rSCCM could inhibit the proliferation of DU145 cells by arresting the cell cycle in the G1 phase in a dose-dependent manner. This growth arresting activity was abolished by boiling rSCCM for 5 min. The G1 growth-inhibiting activity of rSCCM was also detected in other prostate-originated cancer cells examined (i.e., LNCaP and PC-3) but not in other cells (ASC-17D, HepG2, SK-N-SH, and NIH3T3). Western blot analysis of partially purified growth inhibiting fractions with the clusterin antibody showed that the cytostatic factor in rSCCM was not c1usterin. This cytostatic factor was semi purified by DEAE-Sepharose, ammonium sulfate precipitation, and Phenyl-Sepharose column chromatography, and was estimated to have a molecular weight of 88 kDa by Sephacryl S-300 gel filtration.

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활락효영단합인삼양위탕(活絡效靈丹合人蔘養胃湯)이 혈관신생(血管新生) 억제(抑制)에 미치는 영향(影響) (Effect of Whalakyuoleyng-dan plus Yinsamyangwui-tang on Anti-angionesis)

  • 고기완;박준혁;강희;김성훈;유영법;심범상;최승훈;안규석
    • 대한한방종양학회지
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    • 제7권1호
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    • pp.77-97
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    • 2001
  • Anti-angiogenesis is one of therapies which have been high-lightened on the research of cancer treatment. Anti-angiogenesis means that new blood vessels are created from a existing capillary tube and it is a important process on metastasis and permeation when cancer is created or formed. Since angiogenesis have been under research, a complete recovery oriented treatment against cancer have been suggested blocking metastasis, delaying the growth of cancer cell, and blocking the supply of oxygen and nutritive substance through the web of blood vessels. Until now, there are several anti-angiogenesis, which have been known to the public, such as thalidomide, angiostatin, endostatin, 2-methoxyestradiol, TNP-470, and marimastat, etc. Additionally, 17 clinical testing projects about anti-angiogenesis are on the process in NCI(National Cancer Institute). Especially, TNP-470 showed effectiveness against cancer on clinical testing after finishing animal testing. Based on existing researches showing that Yinsamyangwui-tang is effective to strengthening body resistance and Whallakhyolenyng-dan effects cells on the inside of blood vessel because Whallakhyolenyng- dan restrains cell adhesion during the restraining period of a blood vessel, I tried to research the effect of Whalakhyolenyng-dan plus Yinsamyangwui-tang on angiogenesis. I made a conclusion putting into operation through using SK-Hep-1 (KCLB 30052), A549(KCLB 10185), AGS(KCLB 21739), and BCE(Bovine Capillary Endothelial Cell). Followings are the results of my experimental research: 1. According to the researching results of anti-cancer activation against cancer cell, Whallkhyoleyng dan plus Yinsamyangwui-tang decreased the number of cancer cells -- While injecting $600{\mu}g/ml$, injected groups decreased 3.1% more comparing with the contrastive group of SK-Hep-1, 49.7% more comparing with the contrastive group of A549, and 31.0% more comparing with the contrastive group of AGS. 2. According to the researching results of DNA composition effect between BCE and cancer cell, Whallakhyoleyng-dan plus Yinsamyangwui-tang reduced the rate of SK-Hep-1 synthesis inhibition by 59.1% at $600{\mu}g/ml$ intensity comparing with contrastive group; for A549, 72.6%; for AGS, 6.1%, for BCE, 28.9%. 3. According to the researching results about the effect of BCE cell to angiogenesis, angiogenesis was restrained at $400{\mu}g/ml$ intensity during 18 hours observation. 4. In the case of aortic ring assay, the half level of angiogenesis was reduced comparing with the contrastive group while injecting with $400{\mu}g/ml$ intensity; with $800{\mu}g/ml$, under 10% comparing with contrastive group; and with $1600{\mu}g/ml$, complete restrain. According to the above results, Whallakhyoleyng-dan plus Yinsamyangwui-tang was proved to have an anti-angiogenetic effects.

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Sterols Isolated from Nuruk (Rhizopus oryzae KSD-815) Inhibit the Migration of Cancer Cells

  • Lee, Dae-Young;Lee, Sang-Jin;Kwak, Ho-Young;Jung, La-Koon;Heo, Ji-Eun;Hong, Sung-Youl;Kim, Gye-Won;Baek, Nam-In
    • Journal of Microbiology and Biotechnology
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    • 제19권11호
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    • pp.1328-1332
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    • 2009
  • An activity-guided fractionation method was used to isolate anticancer components from Nuruk (Rhizopus oryzae KSD-815:KSD-815). Dried powder of KSD-815 was extracted with 80% methanol and partitioned successively using n-hexane, ethyl acetate, n-butanol, and water. The n-hexane and n-butanol fractions showed a strong antimigratory effect on human cancer cells. Both of these fractions were subjected to separation and purification procedures using silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies to afford four purified compounds. These were identified as ergosterol peroxide (1), stigmast-5-en-$3\beta$,$7\beta$-diol (2), ergosta-7,22-dien-$3\beta$,$5\alpha$,$6\beta$,$9\alpha$-tetraol (3), and daucosterol (4), respectively, by spectroscopic methods such as nuclear magnetic resonance spectrometry, mass spectrometry, and infrared spectroscopy, and comparison with those in the literature. Compounds 1-4 were isolated from KSD-815 for the first time. Compounds 1 and 4 inhibited the migration of MDA-MB-231 cells at concentrations lower than $20\;{\mu}M$.