• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.7 no.1
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• pp.19-37
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• 2001

Object When angiogenesis is excessive, Cancer, RA, Blindness, Psoriasis, Hemangioma, Diabetic retinopathy, Granulation, etc are induced. On the contrary, when it is insufficient, Stroke, Heart disease, Ulcer, Infertility, Scleroderma, artherosclerosis, delay of the wound recovery, etc occur. In recently, the methods which is control of abnormal angiogenesis are researching actively in relathion to anticancer research. This study is search for effective drugs which suppress this angiogenesis, in the ingredients of the herbs that invigorate and dispel blood stasis using to treat intravascular coagulation in the oriental herbal medicine. Methods We maked 80 % methanole extracts of Cnidii Rhizoma, Olibanum, Myrrha, Corydalidis Tuber, Curcumae Radix, Curcumae longe Rhizoma, Zedoariae Rhizoma, Salviae miltiorrhizae Radix, Polygoni cuspidati Rhizoama, Leonuri Herba, Persicae Semen, Carthami Flos, Trogopterorum Faeces, Achyranthis Bidentatae Radix, Manitis Squama, Eupolyphaga, Hirudo, Tabanus, Lycopi Herba, Artemisiae anomalae herba, Vaccariae Semen, Sappan Lignum, Gleditsiae Spina, Draconis Resina, Leonunari Semen, Selaginelliae Folium, Spatholobi Caulis, and these extracts were tested for MTT viabilaty test, BrdU incorporation, Tube foramtion assay on ECV304(immotalized human umbilical vein endothelial cell) at the concentration of $50{\mu}g/ml$, $100{\mu}g/ml$, $200{\mu}g/ml$, $400{\mu}g/ml$ Results All extracts except Draconis Resina have no cytotoxicity at the $100{\mu}g/ml$, and in BrdU incorporation test, proliferation rate were reduced below 60% at the concentaraion of $100{\mu}g/ml$ by Zedoariae Rhizoma, Sappan, Lignum Gleditsiae, Spina Draconis Resina Vaccariae Semen. Zedoariae Rhizoma Sappan Lignum Gleditsiae Spina Draconis Resina Vaccariae Semen Olibanum, Achyranthis Bidentatae Radix showed inhibition effects on tube formation of ECV304 at the concentration of $100{\mu}g/ml$. Conclusion At the concentration of $100{\mu}g/ml$ in which cytotoxicity is not found, Zedoariae Rhizoma, Sappan Lignum, Gleditsiae Spina, Vaccariae Semen showed the inhibition effect on proliferation and tubeformation of ECV304.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.478-484
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• 1996

Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL21 harboring a plasmid pYHB101. The maximum production was 68.7 mg/l when the E. coli strain was cultured at 25$\circ$C for 48 hours in the modified MBL medium containing 10 g/l glucose with 1 mM IPTG induction at 2 hours after inoculation. The rhEGF was purified upto 267 folds by Amberlite XAD- 7 chromatography, ultrafiltration, and DEAE Sepharose fast flow ion exchange chromatography with an overall yield of 66.6%. The purified rhEGF was further separated into two fractions by HPLC. The N-terminal amino acid sequence of the second fraction was Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His. The effect of rhEGF on the DNA synthesis was examined using in vitro biological assay based on the incorporation of 5'-bromo-2'- deoxy-uridine (BrdU). The purified rhEGF shows no difference with natural human epidermal growth factor (nhEGF) in N-terminal amino acids residues and biological activity. From the results, we concluded that rhEGF produced from E. coli harboring the plasmid pYHB101 was apparently the same as nhEGF.

• 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 Korean Traditional Oncology
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• v.11 no.1
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• pp.119-134
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• 2006

Shiquandabutang is very famous prescription for tonifying vital energy. We examined the anti-metatstastic effect of Shiquandabutang with in vitro invasion assay model. We performed the following experiments and the results are listed below:Cell viability assay was carried to determine the dose of Shiquandabutang. At lower dose under 200 ${\mu}g/m{\ell}$ (89.6%) viability was very high. But, viability downed as dose grows. At the dose of 600 ${\mu}g/m{\ell}$ (54.2%) viability was almost half of that of control. And at high dose of 1000 ${\mu}g/m{\ell}$ (15.8%) viability was very pure. In BrdU incorporation assay, Shiquandabutang treated groups showed the decreased DNA synthesis rate compared with control group.(200 ${\mu}g/m{\ell}$ (64.4%), 400 ${\mu}g/m{\ell}$ (7.3%)) The results of gelatinase assay showed that Shiquandabutang decreases the gelatinolytic activity of MMP-9. We examined tube formation assay and the result was that Shiquandabutang ihhibits the tube formation at the dose of 200 ${\mu}g/m{\ell}$ and 400 ${\mu}g/m{\ell}$. We examined rat aortic ring assay and the result was that Shiquandabutang ihhibits the angiogenesis of the rat aortic ring at the dose of 400 ${\mu}g/m{\ell}$. From our research, part of the mechanism underlying anti-metastastic effect of Shiquandabutang was proven in vitro. Moreover, we knew that Shiquandabutang is more effectively inhibits the angiogenesis at high dose.

• Journal of the East Asian Society of Dietary Life
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• v.20 no.1
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• pp.30-36
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• 2010

This study was conducted to investigate the effects of blueberry extract on cell death, ROS and gene expression patterns associated with the anti-cancer activity in human breast cancer MCF7 cells. To accomplish this, 20 mg/mL concentration of blueberry extract was added to the cell culture for 0, 6, 12, 24 or 48 h, after which the effects were evaluated by various analyses. MTT assay showed that the cellular activities decreased rapidly during the first 12 h of treatment. During this period, dual staining with Hoechst33322 and propidium iodide also produced a similar trend in which the dead or dying cells increased sharply. Furthermore, evaluation of BrdU incorporation as an index for cell proliferation revealed a marked decrease during the first 12 h of treatment, suggesting that anticancer activity involves the inhibition of cell proliferation and induces cell death. ROS also increased according to the duration of the treatment, indicating intracellular accumulation is associated with the cell death. RT-PCR analysis revealed significant decreases in anti-apoptotic (Bax) and increases in pro-apoptotic gene expressions (Bci-2, caspase- 3, and 9) (p<0.05). Taken these together, blueberry extract induces ROS accumulation in MCF7 cells, causing inhibition of cell proliferation and eventually leading to cell death. This cell death was associated with apoptotic gene expression in blueberry-treated cells for up to 24 h.