• 생명과학회지
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• 제19권5호
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• pp.633-638
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• 2009

저온 진공 공정으로 건조된 참치를 유기용매로 추출하여 이들 참치 추출물 및 분획물들에 의한 인체 결장암 및 섬유육종세포에 대한 증식 및 세포 내 활성산소종 억제 효과에 대해 검토하였다. 건조 참치의 A+M 추출물과 MeOH 추출물을 0.5, 1, 및 5 mg/ml의 농도로 인체 섬유육종세포(HT1080)에 처리했을 때 농도 의존적으로 인체 암세포 증식을 억제시켰으며(p<0.05), 특히 A+M 추출물이 MeOH 추출물에 비하여 그 억제효과가 우수하였다. 건조 참치 추출물로부터 얻어진 분획물들의 경우 특히 hexane과 85% aq. MeOH 분획물들에 의한 암세포 증식 억제효과가 높았다. 인체 결장암세포(HT-29)에 대한 결과로 A+M 추출물은 인체 섬유육종세포(HT1080)의 결과와 비교했을 때 다소 암세포 증식 억제 효과가 낮았으나 5 mg/ml의 첨가농도에서 95%로 암세포 증식 억제효과를 보였고(p<0.05), MeOH 추출물도 앞서 A+M 추출물 결과와 유사하게 인체 섬유육종세포(HT1080)에 비해 낮은 암세포 성장 억제효과를 보였지만, 첨가농도 5 mg/ml에서는 93%로 높은 암세포 증식 억제효과를 나타내었다(p<0.05). 건조 참치 분획물들의 경우도 앞서의 인체 섬유육종세포(HT1080)와 유사하게 모든 분획물들에서 농도 의존적으로 억제효과가 높은 것을 살펴 볼 수가 있었고, 인체 결장암세포(HT-29)에서도 hexane과 85% aq. MeOH 분획물에 의한 항암효과가 우수하였다. In vitro 항산화실험에서 건조 참치 A+M 추출물 및 MeOH 추출물을 농도별로 인체 섬유육종세포(HT1080)에 처리하였을 때 A+M 추출물(10 mg/ml 농도)를 제외하고 측정기간 120분 동안 모든 추출물들이 blank군과 control군에 비해 세포 내 활성산소종을 크게 감소시켰다. 건조 참치 분획물들 중 BuOH 및 85% aq. MeOH 분획물들은 세포내 활성산소종을 크게 감소시키는 우수한 항산화 효과를 나타내었다.

• Journal of Ginseng Research
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• 제22권3호
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• pp.216-221
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• 1998

We examined the anti-invasive activity of ginsenosides Rhl, Rha on the highly metastatic HT1080 human fibrosarcoma cell line. In vitro invasion assay showed ginsenoside Rhr reduced tumor cell invasion through a reconstituted basement membrane in a transwell chamber more than ginsenoside Rh1. Significant down-regulation of matrix metalloproteinase-9 (MMP-9) by ginsenosides Rh, and Rh2 was detected by Northern blot analysis. However, the expression of MMP-2 was not affected by Rh, and Rhr. The expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) was increased by Rhl after 0.5, 1 or 3 day-treatment but reduced after 6 day-treatment. However, the expression of TIMP-2 was not changed by treatment with Rh2. Plasminogen activator inhibitor (PAI) and urokinase-type plasmlnogen activator (uPA) were not changed by treatment with Rh1 and Rh2 for 3 and 6 days. Quantitative gelatin-based zymography confirmed a markedly reduced expression of MMP-9 but MMP-2 after treatments with ginsenosides Rhl and Rha. These results suggest that down-regulation of MMP-9 contributes to the anti-invasive activity of ginsenosides Rhl and Rhr in the HT1080 cells.

• 생명과학회지
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• 제29권3호
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• pp.287-294
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• 2019

갯메꽃은 해안 사구에 분포하며 식물로 높은 환경 적응력을 가지고 있으며 갯메꽃은 항산화, 해열, 살균, 이뇨작용 등에 효과가 있는 것으로 알려져 있다. 본 연구에서는 암전이 과정에서 중요한 역할을 한다고 알려진 matrix metalloproteinases (MMPs)의 일종인 MMP-2와 MMP-9의 활성에 대한 갯메꽃의 억제효과를 methylene chloride (MC) 및 methanol (MeOH) 추출물과 조추출물, 유기용매 분획물을 시료로 하여 인체 섬유육종 HT-1080 세포를 이용하여 ELISA (Enzyme-linked immunosorbent), gelatin zymography, Wound healing assay, RT-PCR, western blot 실험을 통해 조사하였다. 갯메꽃의 MC 및 MeOH 추출물, 조추출물과 4가지 유기용매 분획물이 HT-1080 세포에서 MMP-2와 MMP-9 생성 억제 활성을 나타냄을 확인하였다. 그 중에서도 n-hexane, 85% aq.MeOH 분획물이 전반적으로 MMP-2와 MMP-9에 대한 높은 억제활성을 보였다. 이들 결과로부터 85% aq.MeOH과 n-hexane 분획물에 MMP 억제 활성이 높은 물질이 존재할 것으로 추측되어지며, 갯메꽃 추출물 및 분획물이 암침윤 및 전이를 억제하는 효과적인 항암소재로서의 가능성이 있음을 제시한다. 이에 본 연구팀은 85% aq.MeOH과 n-hexane 분획물에 있는 활성물질을 분리하기 위한 연구를 추가적으로 수행할 계획이다.

• 동의생리병리학회지
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• 제18권6호
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• pp.1686-1693
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• 2004

Ekong-san(EKS) was expected to have inhibitory effects on angiogenesis, considering the fact that its constituents such as Ginseng Radix, Glycyrrhizae Radix and Citri Pericarpium were reported to inhibit angiogenesis. Moreover, recently several metabolites transformed by the human intestinal microflora were reported to enhance effectiveness compared to their crude drugs. Based on these data, this study was designed to confirm whether the EKS metabolites (EKS-M) can significantly exert the anti-angiogenic and anti-metastatic activites. Hence, with EKS and EKS-M, viability assay, proliferation assay, in vitro tube formation assay, gelatin zymogram assay, in vitro invasion assay were carried out. EKS showed less toxicity in ECV304 and HT1080 cells than EKS-M. EKS-M inhibited the proliferation of HT1080 cells by 30% at 200㎍/㎖ and 42% at 400 ㎍/㎖ respectively. Also, EKS-M degraded the tube network at 200㎍/㎖. EKS and EKS-M inhibited the expression of MMP-9 at 200 and 400㎍/㎖ in HT1080 cells. EKS reduced the invasive activity of HT1080 cells through matrigel coated transfilter atthe concentration of 200㎍/㎖ more effectively than EKS-M. These data suggest that EKS and EKS-M has anti-angiogenic and anti-metastatic activities.

• 대한암한의학회지
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• 제11권1호
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• pp.55-63
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• 2006

This study was conducted to investigate angiogenic inhibitory effect of Zingiberis Rhizoma methanol extract using ECV-304 cells and HT1080 fibrosarcoma cells. The viability of ECV-304 was 30% at 50${\mu}g/m{\ell}$ of Zingiberis extract and that of HT1080 was 30% at 100${\mu}g/m{\ell}$. Using the BrdU incorporation assay, Zingiberis inhibited the DNA synthesis of ECV-304 and HT1080 by 70% and 50 % at 200${\mu}g/m{\ell}$. In tube formation assay, at 10${\mu}g/m{\ell}$ of Zingiberis, tube network began to degrade and at higher doses, it was completely destroyed. Zymography demonstrated that Zingiberis extract decreased MMP-9 at 10${\mu}g/m{\ell}$ and higher doses remarkably inhibited the expression of MMP-9. These data indicate that Zingiberis Rhizoma has angiogenic inhibitory effects and shows the possibility of future anti-metastatic drug.

• Toxicological Research
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• 제14권4호
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• pp.569-575
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• 1998

One of the most frequent dejects in human cancer is the uncontrolled activation of the ms-signaling pathways. Significant evidence has accumulated to directly implicate members of the matrix metalloproteinases (MMPs) in tumor invasion and metastasis formation. We have previously shown that MMP-9 expression was significantly enhanced in the ras-tranfected HT1080 human fibrosarcoma cells at the mRNA level. In the present study, we investigated the roles of MMP-2 and -9 on the H-ras-induced invasive phenotypes of MCF 10A human breast epithelial cells and HT 1080 human fibrosarcoma cells. We show that H-ras is able to induce or enhance a signaling pathway leading to the enhancement of an invasive phenotype in both MCF10A and HT1080 cells as determined by matrigel invasion assay. We then examined the effect of H-ras activation on the expression of MMP-2 and -9 by measuring enzymatic activities and mRNA levels. Our data clearly demonstrated that H-ras prominently induces expression of MMP-2 in MCF10A cells, while it efficiently up regulates MMP-9 in HT1080 cells. Taken together, these findings suggest that the correlation between ras-mediated invasiveness and enhanced expression of MMPs may be cell type-specific: MMP-9 is closely associated with the invasive phenotype induced by ras activation in fibrosarcoma cells, whereas MMP-2 is more likely associated with it in epithelial cells.

• KSBB Journal
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• 제29권5호
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• pp.343-352
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• 2014

In the present study, antioxidant activities of two crude pigments (acetone and MeOH) and their solvent fractions (n-hexane, 85% aq.MeOH, n-BuOH, and water fractions) from red crab shell were evaluated by measuring 1,1-diphenyl-2-picryl hydrazyl (DPPH), peroxynitrites, and degree of production of reactive oxygen species (ROS) in HT 1080 cells as well as the extent of oxidative damage of genomic DNA purified from HT 1080 cells. From comparative analysis, 85% aq.MeOH fraction showed the strongest scavenging effect on both peroxynitrite in vitro and intracellular ROS in HT 1080 cells. Protective activities of these samples against hydroxyl radical-mediated genomic DNA damage were also investigated. 85% aq.MeOH and n-BuOH fractions significantly inhibited oxidative damage of purified genomic DNA. On the other hand, we investigated their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. All samples significantly reduced NO production. Among the samples, n-hexane and water solvent fractions most effectively inhibited NO.

• Animal cells and systems
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• 제13권2호
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• pp.113-118
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• 2009

Extracts derived from various medical mushrooms have been reported to have antitumor and immuno-modulatory properties. In order to investigate the antitumor activity of keumsa Linteusan, the water extract of Phellinus Iimteus, HT1080 cells, a human fibrosarcoma cell line, were treated with it and changes in cellular migration potential was tested in vitro. At a concentration range below 1,000 $\mu$g/mL, Linteusan blocked, in a dose dependent manner, the migration of cells through Matrigel as well as Boyden chamber without affecting the viability of the cells. Prolonged treatment of HT1080 cells with Linteusan suppressed TNFa induced production of matrix metalloproteinase (MMP)-9 as well as basal level expression of MMP-2. Linteusan also affected the adhesion of the cells to fibronectin-coated surfaces. The effect of Linteusan on cell signaling pathways was also tested. Linteusan specifically affected TNF-$\alpha$ induced phosphorylation of AKT in a dose-dependent manner, while phosphorylation levels of ERK remained unaffected. These data indicate that Linteusan blocks the migration of HT1080 cells by affecting various processes associated with cell migration such as the expression of matrix degradingenzymes,cell adhesion, and AKT-medicated cellular signaling pathways.

• BMB Reports
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• 제29권5호
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• pp.484-487
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• 1996

The matrix metalloproteinases (MMPs) are members of a unique family of proteolytic enzymes that degrade components of the extracellular matrix. Significant evidence has accumulated to directly implicate members of the MMPs in tumor invasion and metastasis formation. To investigate the correlation between ras oncogene and MMP gene expression in various tumor cells, we detected mRNAs for the ras, MMP-2 and MMP-9 (72 kD and 92 kD type IV collagenases, respectively) genes in nine human tumor cell lines. The ras gene was expressed in seven cell lines; MMP-2 in three; MMP-9 in two cell lines tested. There was no direct correlation between the ras oncogene and MMP expression. A clear difference in the mRNA expression between MMP-2 and MMP-9 was observed among the cell lines. As an approach to study the effect of the ras oncogene on metastasis, we examined the expressions of MMP-2 and MMP-9 in HT1080 cells transfected with the v-H-ras gene. MMP-9 expression was Significantly enhanced in the ras-transfected HT1080 cells compared with the nontransfectants while ras transfection did not affect the expression of MMP-2. These results suggest the possible inducing effect of the ras oncogene on the metastasis by activation of the MMP-9 gene in HT1080.

• 동의생리병리학회지
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• 제17권2호
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• pp.525-528
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• 2003

Spatholobus Suberectus dunn. has been applied to blood stasis in oriental medicine. We selected one potent ethyl acetate subfraction-2 from Spatholobus Suberectus Dunn.(SSD) from anti-metastasis screening. It exerted the cytotoxicity against HT1080 and B16BL6 with the IC50 of 60 ug/ml and also significantly inhibited tumor cell induced platelet aggregation (TCIPA). It effectively didn't inhibit cell adhesion of HT1080 to matrigel coated wells, while it inhibited the cell invasion of HT1080 at the doses of 10, 20, 40 μg/ml by Boyden chamber assay. It effectively suppressed lung metastasis by B16BL6 melanoma in C57BL6 mice. These results indicate that the EtOAc subfraction-2 of Spatholobus Suberectus Dunn. can be applied to caner treatment with anti-metastatic activity.