• Journal of Korean Traditional Oncology
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• v.11 no.1
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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.

• Journal of Life Science
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• v.28 no.3
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• pp.293-299
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• 2018

Viticis fructus (fruits of Vitex rotundifolia) is the dried fruit from Vitex rotundifolia; is a traditional medicine for treating inflammation, migraines, chronic bronchitis, headaches, eye pain, and gastrointestinal infections; and demonstrates various bioactivities, including anti-allergic, anti-cancer, and anti-inflammatory effects, which are partly due to its phenolic compound content. This study examines the inhibitory effects of viticis fructus (fruits of Vitex rotundifolia) on MMP-2 and MMP-9 expression using gelatin zymography and RT-PCR in phorbol-12-myristate-13-acetate (PMA)-induced HT-1080 fibro-sarcoma cells. Fruits of Vitex rotundifolia were extracted twice using dichloromethane ($CH_2Cl_2$) and methanol (MeOH). The combined crude extracts ($CH_2Cl_2$ and MeOH) significantly inhibited MMP-2 and MMP-9 activities in gelatin zymography. The combined extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq. MeOH), n-butanol, and water, successively according to polarity. Among all solvent-partitioned fractions, 85% aq. MeOH fractions showed the strongest inhibition on the activation of MMP-2 and MMP-9 in gelatin zymography. In PMA-stimulated HT-1080 cells, the expression levels of MMP-2 and MMP-9 mRNA were also greatly inhibited by the 85% aq. MeOH fraction. These results suggest that viticis fructus can be used as an excellent source for anti-invasive agents.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.277-288
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• 2003

It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HTl080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP , suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

• Journal of the Korean Society of Food Culture
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• v.24 no.6
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• pp.749-755
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• 2009

We investigated the chemical components of red onion powder dried using the low temperature vacuum method and the inhibitory effects of solvent extracts of the dried red onion powder on the growth of HT-1080 human fibrosarcoma and HT-29 human colon cancer cells and $H_2O_2$-induced oxidative stress. The moisture content of the dried red onion powder was 17.95%, while the vitamin C content was 96 mg/100 g and the total phenols content was 39.1 mg/mL. The inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts of the red onion powder on the growth of HT-1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was greater on the growth of HT-29 cells, while the A+M extracts had a higher inhibitory effect than the MeOH extracts. Treatment with the hexane, 85% aq. methanol, butanol and water fractions of the extract led to significant inhibition of the growth of both cancer cell lines (p<0.05). Among the fractions, the hexane and 85% aq. methanol fractions showed a greater inhibitory effect. To determine the protective effect on $H_2O_2$-induced oxidative stress, a DCFH-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including the crude extracts of dried red onion, appeared to lead to a significant reduction in the levels of intracellular reactive oxygen species (ROS), and these reductions occurred in a dose dependent fashion (p<0.05). Among the fractions, the 85% methanol fraction showed the greatest protective effect on the production of lipid peroxides.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.4 no.1
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• pp.131-146
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• 1998

To examine the effect of Shiquandabutang on the metastasis of cancer, the following experiments were made. Before the main experiments, the cytotoxicity was measured by putting Shiquandabutang sample in HT1080. Then zymography was made to examine the change of gelatinolytic activity. And western blotting was carried out to examine the changes of Fos, Jun, Ets, the transcription factors of MMP-2, MMP-9, and Erk, JNK on signal transduction pathway to AP-1. Third, in vitro invasion assay with transwells coated by collagen and matrigel was carried out. From the results of the above the following conclusions were obtained. 1. The experimental result about cytotoxicity of Shiquandabutang against HT1080 was as below. The stained cell count after being treated by Shiquandabutang sample $400{\mu}g/ml$ for 24 hours was 0.9% of total cells, and the stained cell count by Shiquandabutang sample $100{\mu}g/ml$ was 1.5% of total cells. Both were near the level of control group which showed 0.6% stained. 2. The result of collagenase assay was as below. In Shiquandabutang sample $400{\mu}g/ml$, MMP-2 was reduced as compared with TPA control group, and the band of MMP-9 induced by TPA disappeared. In Shiquandabutang sample $800{\mu}g/ml$, both bands of MMP-2 and MMP-9 disappeared. 3. The results of western blots for Jun, Fos, Ets, Erk, JNK were as below. In Shiquandabutang sample $200{\mu}g/ml$, Ets was reduced, and Fos were increased. 4. The result of invasion assay was as below. The number of cells which migrated across transwell membrane in Shiquandabutang-treated group was less than that of +TPA control group. From the above results, it was concluded that Shiquandabutang might control the appearing and acting of collagenase not by the MMP-2, -9 promoter but by other way.

• Journal of the Korean Applied Science and Technology
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• v.41 no.2
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• pp.447-458
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• 2024

In this study, the antioxidizing effect of 2,3-dicaffeoyl-epi-quinic acid (DEQA) was investigated. The antioxidant activity was evaluated by measuring the scavenging effect on DPPH radical and peroxynitrite and the reducing power on ferric ion. DEQA showed a scavenging effect and reducing power comparable to vitamin C used as a positive control. Also, DEQA effectively inhibited production of intracellular reactive oxygen species (ROS) in HT-1080 cells, showing the scavenging ratio of 43.8% even at 10 µM concentration of DEQA after 2 hours in HT-1080 treated with H₂O₂. In addition to this, DEQA inhibited the production of nitric oxide (NO) very effectively in Raw 264.7 cells. The above results suggest that DEQA has the potential to be developed as a natural antioxidant.

• KSBB Journal
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• v.24 no.2
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• pp.201-206
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• 2009

Whole plants of Corydalis heterocarpa were extracted twice with $CH_2Cl_2$ and MeOH in turn. The combined crude extracts were concentrated in vacuo and then partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was fractionated with n-hexane and 85% aq. MeOH, and the aqueous fraction was also further fractionated with n-BuOH and $H_2O$, successively. Growth inhibition effects of crude extracts and their solvent fractions were evaluated in AGS, HT1080, U-937, MCF-7 and HT-29 human cancer cells using MTT assay. The inhibitory effects of solvent fractions were increased in a dose-dependent manner. Among these tested samples, 85% aq. MeOH fraction showed the most potent inhibitory effect on the growth of human cancer cells. These results suggest that active compounds having much stronger anticancer effect can be isolated from Corydalis heterocarpa.

• The Korean Journal of Food And Nutrition
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• v.32 no.3
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• pp.208-215
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• 2019

In this paper, we investigate to determine quality characteristics, fatty acid composition and cytotoxic effect of extracts and fractions from whole Lycopus lucidus Turcz. roots. Additionally, we evaluated cytotoxic activity against the growth of human fibrosarcoma cells (HT-1080) and human gastric adenocarcinoma (AGS), human colon cancer cell (HT-29) lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Acetone+methylene chloride (A+M) and methanol (MeOH) extracts from L. lucidus Turcz. were obtained through solvent extraction. Then we further fractionated both extracts with n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water. In fatty acid composition, L. lucidus Turcz. contained 33.2% of 18:1n-9 and 1.81% of 18:3n-3, respectively. The incorporation of treatment with A+M and MeOH extracts and n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water fractions dose-dependently increased cytotoxicity against the growth of HT-1080 and AGS, HT-29 cancer cells (p<0.05). The A+M extract had a higher inhibitory effect on the growth of all cancer cells in comparison to MeOH extract. Among the fractions, the 85% aq. MeOH and n-hexane fractions showed a higher inhibitory effect after proliferating the three cancer cells. These results suggest that the 85% aq. MeOH and n-hexane fractions have a potential to inhibit the growth of human cancer cell lines.

• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.180-186
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• 2009

Whole plants of Vitex rotundifolia were extracted for 2 days with methylene chloride ($CH_2Cl_2$) followed by extraction of the residue for an additional 2 days. The same procedure was also applied using methanol (MeOH). The two crude extracts were combined and partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was further partitioned between n-hexane and 85% aq. MeOH, and the aqueous layer was also further fractionated with n-BuOH and $H_2O$, successively. From the 85% aq. MeOH fraction, one compound was isolated through the repeated HPLC. According to the results of physicochemical data including NMR and MS, the chemical structure of the compound was determined as artemetin (1). The antiproliferative effects of the crude extracts, fractions, and compound against HT1080, AGS, MCF-7 and HT-29 human cancer cells were compared with the control by using MTT assay. In the comparative analysis, the 85% aq. MeOH fraction exhibited the strongest antiproliferative effects on human cancer cell lines in a dose-dependent manner (p<0.05). In addition, exposure of compound 1 isolated from 85% aq. MeOH fraction led to strong antiproliferative effect in HT1080 cancer cell lines. These results suggest that the extracts and compound isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents.

• Journal of Korean Traditional Oncology
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• v.11 no.1
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• pp.41-54
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• 2006

Ekongsan (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 ${\mu}g/m{\ell}$ and 42% at 400 ${\mu}g/m{\ell}$ respectively. Also, EKS-M degraded the tube network at 200 ${\mu}g/m{\ell}$. EKS and EKS-M inhibited the expression of MMP-9 at 200 and 400 ${\mu}g/m{\ell}$in HT1080 cells. EKS reduced the invasive activity of HT1080 cells through matrigel coated transfilter at the concentration of 200 ${\mu}g/m{\ell}$ more effectively than EKS-M. These data suggest that EKS and EKS-M has anti-angiogenic and anti-metastatic activities.