中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (1): 49-51.

• 论著 • 上一篇    下一篇

甲磺酸伊马替尼对人黑素瘤M14细胞系凋亡的影响

高莹,陈浩,关杨,陈佳,曾学思,孙建方   

  1. 南京 中国医学科学院北京协和医学院皮肤病研究所
  • 收稿日期:2009-03-09 修回日期:2009-05-07 出版日期:2010-01-15 发布日期:2010-01-05
  • 通讯作者: 高莹 E-mail:srxying@sina.com
  • 基金资助:

    中国医学科学院青年基金资助（IMBF20060602）；江苏省六大高峰人才基金（06-B-72）

Effects of imatinib mesylate on the apoptosis in human melanoma cell line M14

2, 2, 2, 2   

  • Received:2009-03-09 Revised:2009-05-07 Online:2010-01-15 Published:2010-01-05

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摘要：

目的 探讨甲磺酸伊马替尼对体外培养人黑素瘤M14细胞凋亡的影响。方法 Annexin-V/PI法和TUNEL法观察5 μmol/L、10 μmol/L和20 μmol/L三种浓度甲磺酸伊马替尼对M14细胞凋亡的诱导情况，流式细胞仪检测细胞周期和凋亡率变化，DAPI染色观察细胞凋亡形态变化，Western印迹分析检测bcl-2、bax蛋白的表达。结果 三种浓度甲磺酸伊马替尼均能不同程度的诱导M14细胞凋亡，细胞周期显示S期细胞和凋亡细胞比例高于和对照组（P ＜ 0.05），G2/M期细胞比例下降（P ＜ 0.01）。Annexin-V/PI法检测示甲磺酸伊马替尼组早期凋亡率（Q4%）的升高（P ＜ 0.01）。TUNEL法检测凋亡阳性细胞的数目高于对照组（P ＜ 0.01）。经20 μmol/L甲磺酸伊马替尼作用后DAPI染色示凋亡细胞形态改变，同时Western印迹分析示bcl-2表达上调和bax表达下调。结论 甲磺酸伊马替尼能影响细胞周期进程，诱导M14细胞凋亡，可能与活化线粒体凋亡途径有关。

关键词: 黑色素瘤, 甲磺酸伊马替尼, 凋亡

Abstract:

Objective To study the effects of imatinib mesylate on the apoptosis in human melanoma cell line M14. Methods M14 cells were cultured in vitro in the presence of imatinib mesylate at three concentrations （5, 10 and 20 μmol/L） for 96 hours. Sebsequently, annexin V-FITC and propidium iodide （PI） double staining flow cytometry and terminal deoxynucleotidyl transferase （TdT）-mediated dUTP nick end labeling （TUNEL） were used to detect the cell cycle and apoptosis, respectively, DAPI staining to observe the morphological changes, and Western blot to measure the protein expressions of bcl-2 and bax in cells. Results Imatinib mesylate of the three concentrations could induce an evident increase in the apoptosis in M14 cells. Compared with untreated M14 cells, an increase of cell population in S phase was observed in imatinib mesylate-treated cells （P ＜ 0.05）, along with a decline in cell population in G2/M phases （P ＜ 0.01）. Annexin V/PI double staining and TUNEL revealed a significant increase in the rate of early apoptosis and in the acount of apoptotic cells, respectively, in M14 cells treated with imatinib mesylate of the three concentrations （all P ＜ 0.01）. After treated with imatinib mesylate of 20 μmol/L, there was a morphological change characteristic of apoptosis in M14 cells, together with an upregulated expression of bcl-2 （t = 15.46, P ＜ 0.01） and downregulated expression of bax （t = 25.53, P ＜ 0.01）. Conclusions Imatinib mesylate can interfere with the process of cell cycle of and induce the apoptosis in M14 cells, which may be mediated through mitochondrial pathway.

Key words: Melanoma, Imatinib mesylate, Apoptosis