過酸化水素抵抗性がん細胞における過酸化水素抵抗メカニズムの解析

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  • 長澤 大成
    鹿児島大学大学院 医歯学総合研究科 生体機能制御学講座歯科応用薬理学分野
  • 富田 和男
    鹿児島大学大学院 医歯学総合研究科 生体機能制御学講座歯科応用薬理学分野 兵庫医療大学薬学部医療薬学科　薬理学
  • 桑原 義和
    鹿児島大学大学院 医歯学総合研究科 生体機能制御学講座歯科応用薬理学分野 東北医科薬科大学医学部放射線基礎医学
  • 五十嵐 健人
    鹿児島大学大学院 医歯学総合研究科 生体機能制御学講座歯科応用薬理学分野
  • 髙 裕子
    鹿児島大学大学院 医歯学総合研究科 生体機能制御学講座歯科応用薬理学分野 鹿児島大学大学院 医歯学総合研究科歯科保存学分野
  • 田中 康一
    兵庫医療大学薬学部医療薬学科　薬理学 兵庫医科大学医学部薬理学
  • 北中 純一
    兵庫医科大学医学部薬理学
  • 北中 順惠
    兵庫医科大学医学部薬理学
  • 竹村 基彦
    兵庫医科大学医学部薬理学
  • 西山 信好
    兵庫医療大学薬学部医療薬学科　薬理学
  • 栗政 明弘
    東北医科薬科大学医学部放射線基礎医学
  • 佐藤 友昭
    鹿児島大学大学院 医歯学総合研究科 生体機能制御学講座歯科応用薬理学分野

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  • Analysis of hydrogen peroxide resistance mechanism on hydrogen peroxide resistant cancer cells.

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<p>&lt;purpose&gt;</p><p>Hydrogen peroxide is known as one of ROS which gives oxidative stress to cells and induces apoptosis. However, details of the mechanism for cancer cells by hydrogen peroxide is still unknown.We have established "hydrogen peroxide resistant (HR) cancer cells" that are resistant to high concentration hydrogen peroxide. The mechanism of resistance to hydrogen peroxide acquired by HR cancer cells has not yet been elucidated, however if the mechanism becomes clear, it could be applied to cancer treatment. In this study, therefore, we aimed to elucidate its hydrogen peroxide resistance mechanism and carried out the following experiment.</p><p>&lt;method&gt;</p><p>Cell lines that continued to survive against graded hydrogen peroxide treatment of HeLa (up to 70 μM) and SAS (up to 35 μM) were subjected to hydrogen peroxide at the concentrations of 0, 25, 50, 75 and 100 μM respectively, and the cell viability was examined by WST assay. Subsequently, the endogenous catalase enzymatic activity of HR cancer cells was measured using Catalase Assey Kit (SIGMA).Furthermore, lipid peroxidation of HR cancer cells was analyzed by immunofluorescence using 4-hydroxynonenal (HNE) and 5-lipoxygenase (5-LOX) antibody. HNE is typical lipid peroxidation marker and 5-LOX is known as lipid peroxidase. </p><p>&lt;results and discussion&gt;</p><p>In HeLa and SAS parental cells, they survived to the extent of 25 μM by hydrogen peroxide treatment. On the other hand, stepwise hydrogen peroxide-treated cells survived up to100 μM (HeLa) and 50 μM (SAS), showingresistance to hydrogen peroxide. Analysis of catalase enzyme activity showed significant increase in HeLa HR cells compared with the HeLa parent, but there were no significant differences in SAS cells.</p><p>Furthermore, HNE and 5-LOX expression levels in HR cells were significantly decreased compared with the parental cells byimmunofluorescent staining. </p><p>As mentioned above, the membrane lipid peroxidation is regulated by the expression of 5-LOXrather than a catalase activity. The decreased 5-LOX expression may suppress lipids peroxidation in plasma membrane and leadto retention viability to hydrogen peroxide in HR cells.</p>

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