日本口腔外科学会雑誌
Online ISSN : 2186-1579
Print ISSN : 0021-5163
ISSN-L : 0021-5163
上皮成長因子EGFが退縮型ラット癌細胞ER-1の浸潤, 転移能に及ぼす影響
第3報, EGF長期間処理による酸化ストレスと遺伝子不安定性の誘導
中田 大地柴田 敏之永易 裕樹河野 峰北所 弘行青山 哲也小林 徳栄有末 眞
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2000 年 46 巻 9 号 p. 511-518

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We have previously reported that long-term epidermal growth factor (EGF) treatment enhanced the malignant phenotype of regressor ER-1 cells and might affect the genomic alteration of ER-1 cells. In this study, we examined whether EGF stimulation might induce reactive oxygen species (ROS) in ER-1 cells and whether ROS could affect genomic instability. ROS of ER-1 cells induced by EGF stimulation for 1 month was observed with a confocal laser-scan microscope after 2', 7'-dichlorofluorescein diacetate (DCFH-DA) staining. ROS of ER-1 cells was significantly augmented by EGF stimulation, and ROS spread over the nuclei after stimulation. To study the effect of ROS on ER-1 cells, we measured the amount of glutathione peroxidase (Gpx), radical scavenger enzyme, and 8-hydroxydeoxyguanosine (8-OHdG), which is a mutation indicator. The amount of Gpx was reduced by EGF stimulation and recovered after sodium selenite treatment. Furthermore, to study modulation of the malignant phenotype of ER-1 cells, each treated ER-1 cell was intrapenitorially transplanted into syngenic SHR-rats. Tumorigenicity of ER-1 cells was significantly enhanced by EGF, and selenite treatment completely inhibited the augmentation of tumorigenicity caused by EGF stimulation. This observation was consistent with the in vitro results. Then, to examine the effects of ROS induced by EGF stimulation on the genomic instability of ER-1 cells, DNA fingerprinting assay was done. In this assay, abnormal bands were observed in four clones (4/19 clones) after EGF stimulation. These results suggest that long-term EGF stimulation can affect the genomic alteration of ER-1 cells and that this phenomenon is possibly caused by EGF inducing ROS.

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