中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (9): 633-636.

• 论著 • 上一篇    下一篇

Akt/mTOR活化抗UVB诱导HaCaT细胞凋亡的研究

李金超1,许爱娥2,宋秀祖3,关翠萍3,洪为松4   

  1. 1. 安徽医科大学杭州临床学院
    2. 安徽医科大学附属杭州市第三人民医院皮肤科
    3. 杭州市第三人民医院皮肤科
    4. 杭州市第三人民医院
  • 收稿日期:2009-11-16 修回日期:2010-05-19 发布日期:2010-09-10
  • 通讯作者: 许爱娥 E-mail:xuaiehz@msn.com
  • 基金资助:

    CpG-ODN经Akt/mTOR抗UVB诱导的角质形成细胞凋亡的保护机制研究

Activation of the Akt/mTOR pathway confers protection against UVB-induced apoptosis in HaCaT cells

  • Received:2009-11-16 Revised:2010-05-19 Published:2010-09-10

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摘要：

目的 探讨Akt/mTOR信号通路活化抗中波紫外线（UVB）诱导的HaCaT细胞凋亡。方法 UVB照射角质形成细胞，Western印迹检测Akt/mTOR通路中相关信号分子的动态水平变化。免疫荧光 Hoechst 33342染色观察HaCaT细胞凋亡率。结果 UVB能活化Akt/mTOR信号通路，并在一定范围内（5 ～ 30 mJ/cm2）成剂量依赖性，在一定范围内（5 ～ 30 min）成时间依赖性。EGFR抑制剂PD 153035、PI3K抑制剂LY 294002和mTOR抑制剂雷帕霉素能显著抑制UVB对Akt/mTOR信号通路的活化作用。UVB照射前加入雷帕霉素、LY 294002预处理，HaCaT细胞凋亡率增加。结论 Akt/mTOR活化抗UVB诱导的HaCaT细胞凋亡。

关键词: 紫外线, 角蛋白细胞, 信号转导

Abstract:

Objective To investigate the role of activation of Akt/mTOR pathway in denfense against UVB-induced apoptosis in cultured human skin keratinocyte cell line HaCaT. Methods HaCaT cells were irradiated with UVB at different doses for various durations. Western blotting was performed to detect dynamic changes of Akt/mTOR pathway-related signaling molecule, such as phosphorylated-epidermal growth factor receptor （EGFR）, -Akt, -4EBP1, etc; apoptosis was estimated by staining with DNA dye Hoechst 33342. To evaluate the role of signaling molecules in defense against UVB-induced apoptosis, HaCaT cells were pretreated before irradiation with EGFR inhibitor （PD153035）, PI3K inhibitor （LY294002）, mTOR inhibitor （rapamycin） followed by the detection of expressions of signaling molecule and apoptosis. Results UVB could activate Akt/mTOR pathway in a dose- （5 ~ 30 mJ/cm2） and time- （5 ~ 30 min） dependent manner. PD153035, LY 294002 and rapamycin could inhibit UVB-induced activation of the Akt/mTOR pathway. The apoptosis rate in HaCaT cells was upregulated by pretreatment with rapamycin and LY294002. Conclusion The activation of Akt/mTOR signaling pathway could inhibit the UVB-induced apoptosis in cultured HaCaT cells.

Key words: Ultraviolet rays, Keratinocytes, Signal transduction