南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (04): 375-378.

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过氧化氢抑制三氧化二砷诱导的伯基特氏淋巴瘤细胞凋亡

陆地1, 白晓春1, 刘斌2, 李秀梅1, 邓凡1, 李明1, 程宝鸾1, 罗深秋1   

  1. 1. 第一军医大学细胞生物学教研室, 广东, 广州, 510515;
    2. 第一军医大学南方医院脊柱骨科, 广东, 广州, 510515
  • 出版日期:2004-04-20 发布日期:2004-04-20
  • 基金资助:
    收稿日期:2003-12-5。
    基金项目:国家自然科学基金(39870381和30300397);广东省自然科学基金(980213)
    作者简介:陆地(1969-),男,1997年毕业于昆明医学院,讲师,博士研究生,E-mail:ludi2004@hotmail.com
    通讯作者:罗深秋,男,第一军医大学细胞生物学教研室,教授,博士生导师,电话:020-61648208,E-mail:Luoshq888@163.com

Hydrogen peroxide inhibits arsenic trioxide-induced apoptosis of Burkitt lymphoma cells

LU Di1, BAI Xiao-chun1, LIU Bin2, LI Xiu-mei1, DENG Fan1, LI Ming1, CHENG Bao-luan1, LUO Shen-qiu1   

  1. 1. 第一军医大学细胞生物学教研室, 广东, 广州, 510515;
    2. 第一军医大学南方医院脊柱骨科, 广东, 广州, 510515
  • Online:2004-04-20 Published:2004-04-20

摘要: 目的 探讨过氧化氢(H2O2)对三氧化二砷(As2O3)诱导人类伯基特氏淋巴瘤细胞凋亡的影响及双染法荧光显微技术在检测细胞凋亡中的作用。方法 应用As2O3诱导人类伯基特氏淋巴瘤细胞株BJAB细胞凋亡,采用Hoechst 33342和碘化丙啶(PI)核染料双染法荧光显微镜和透射电镜对细胞死亡进行观察及定量分析。结果 BJAB细胞经As2O3处理后,荧光显微镜下可见大多数细胞出现细胞染色质凝集、核边集、核碎裂等凋亡的特征性改变;在低剂量(200 μmol/L)H2O2的影响下,抑制了As2O3诱导的细胞凋亡,降低细胞的死亡率;细胞的死亡方式由凋亡转变为核固缩性坏死,表现为既无凋亡的特征性改变,又非典型的细胞坏死,死亡细胞的细胞核呈固缩状态而非肿胀。结论 低浓度(200 μmol/L)H2O2可抑制临床治疗浓度的As2O3诱导的人类伯基特氏淋巴瘤BJAB细胞凋亡;双染法荧光显微镜技术不但可明确区分细胞凋亡及坏死,还可判断早期凋亡细胞和晚期凋亡细胞,并可进行定量分析,是一种良好的细胞凋亡检测方法。

Abstract: Objective To examine the effects of hydrogen peroxide (H2O2) on arsenic trioxide (As2O3)-induced apoptosis of human Burkitt lymphoma cells and evaluate the value of fluorescence microscope in analyzing cellular apoptosis. Methods As2O3 was used to induce apoptosis in human Burkitt lymphoma cells BJAB, and the cellular changes were analyzed quanti- tatively using transmission electron microscope and fluorescence microscope after staining with Hoechst 33342/propidium iodide (PI). Results Under fluorescence microscope and electron microscope, BJAB cells displayed chromatin aggregation, nuclear margination and fragmentation in response to As2O3 treatment. In the presence of relatively low levels of H2O2 (200 μmol/L), the cell death resulting from apoptosis was inhibited with pyknosis and necrosis becoming the major pathway, while As2O3 failed to induce cell apoptosis. The cells showed none of the typical markers of apoptosis nor any characteristic necrotic changes, and the nuclei exhibited condensation instead of swelling. In addition, the rate of cell death induced by As2O3 was decreased in the presence of H2O2. Conclusions Low levels of H2O2 (200 μmol/L) inhibits As2O3-induced (at clinically achievable concentrations) apoptosis of the human malignant lymphoma cells. Fluorescence microscopy makes possible quantitative analysis of apoptosis, capable of distinguishing not only cell apoptosis from necrosis, but also membrane-intact from membrane-permeable apoptotic cells.

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