中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (6): 457-459.

• 论著 • 上一篇    下一篇

亚砷酸对角质形成细胞增殖及凋亡的影响

毕新岭, 顾军, 陈志强, 米庆胜   

  1. 1. 第二军医大学长海医院皮肤科;
    2. 中国医学科学院皮肤病研究所;
    3. 泰山医学院皮肤性病科
  • 收稿日期:2001-11-06 出版日期:2002-12-15 发布日期:2002-12-15
  • 通讯作者: 顾军,200433上海,第二军医大学长海医院皮肤科
  • 基金资助:
    2001年卫生部科技专项基金

Effects of Trivalent Arsenicals on Cell Proliferation and Apoptosis in Human E pidermal Keratinocytes

BI Xinling, GU Jun, CHEN Zhiqiang, MI Qingsheng   

  1. Department of Dermatology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
  • Received:2001-11-06 Online:2002-12-15 Published:2002-12-15

PDF (PC)

203

赞

摘要： 目的 研究亚砷酸对不同角质形成细胞增殖及凋亡的影响.方法 以体外培养的角质形成细胞株为对象,不同浓度的亚砷酸处理细胞后,用MTT比色分析法反映细胞增殖变化,用光镜和电镜观察细胞形态,流式细胞仪测定细胞周期分布及凋亡峰的出现,Annexin-V染色荧光照相证实凋亡的发生.结果 0.5～10μmol/L亚砷酸对良性角质形成细胞株HaCaT细胞有明显抑制作用,并呈时间和剂量依赖关系,而该浓度砷剂对A431细胞无明显影响.光镜及HE染色后观察均显示随浓度升高、时间延长,凋亡细胞增多.高于上述范围HaCaT细胞变性死亡增多,流式细胞仪检测细胞周期分布变化,G2M期细胞比例及亚G1期凋亡峰明显升高,Annexin-V法显示有绿色荧光的阳性细胞.而A431细胞无明显形态和细胞周期分布变化.结论 一定浓度的亚砷酸可抑制HaCaT细胞增殖,并具有诱导凋亡作用,对皮肤鳞状细胞癌A431细胞无明显影响,提示良性表皮角质形成细胞株HaCaT对亚砷酸的处理比鳞状细胞癌A431细胞株更敏感.

关键词: 脱噬作用, 亚砷酸盐类, 角蛋白细胞

Abstract: Objective To investigate the effects of trivalent arsenicals on cell proliferation and induction of apoptosis in human epidermal keratinocytes. Methods Human benign epidermal keratinocytes (cell line HaCaT), human epidermal carcinoma cells(cell line A431) were cultured. After treatment with arsenous acid, inhibition of cellular growth was determined by measuring MTT dye absorption of living cells.Apoptosis was assessed with respect to morphological changes by light and electron microscopy and to cell cycle distribution by flow cytometry. An nexin-V binding assay was used to detect the early stage of apoptosis. Results With concentrations ranging from 0.5 to 10 靘ol/L, arsenous acid significantly inhibited the proliferation of HaCaT cells in a dose-and time-dependent manner. By light and electron microscopy, morphological changes revealed characteristics of apoptosis. But A431 cells showed no obvious change. DNA flow cytometric an alysis indicated that arsenousacid induced an arrest in G2M phase and sub-G1p hase in HaCaT compared with A431 cells. The green flurorescence indicated early stage of apoptosis in HaCaT cells by annexin-V binding assay. Conclusion Arsenous acid may inhibit the proliferation of HaCaT cells and induce apoptosis, but does not affect A431 cell line obviously, which suggests that HaCaT cells are more sensitive to arsenous acid compared with A431 cells.

Key words: Arsenites, Keratinocytes, Apoptosis