中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (6): 418-421.

• 论著 • 上一篇    下一篇

5-氮-2′-脱氧胞苷对黑素瘤细胞IGFBP7 表达及细胞增殖的影响

薛燕宁1,陈浩1,周武庆2,宋亚丽3,曾学思1,孙建方1   

  1. 1. 南京 中国医学科学院北京协和医学院皮肤病研究所
    2. 南京医科院皮研所
    3. 山东大学附属省立医院皮肤科
  • 收稿日期:2009-12-09 修回日期:2010-03-17 出版日期:2010-06-15 发布日期:2010-06-08
  • 通讯作者: 薛燕宁 E-mail:xyning52@163.com
  • 基金资助:

    国家自然科学基金

Effects of 5-aza-dc on the expression of insulin-like growth factor binding protein 7 （IGFBP7） in and proliferation of melanoma cells

  • Received:2009-12-09 Revised:2010-03-17 Online:2010-06-15 Published:2010-06-08

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摘要：

目的 探讨去甲基化药物5-氮-2′-脱氧胞苷对体外培养黑素瘤细胞系A375和M14细胞胰岛素样生长因子结合蛋白7（IGFBP7）的表达及细胞增殖的影响。方法 体外培养黑素瘤A375和M14细胞，采用RT-PCR和免疫细胞化学法分别检测10 μmol/L 5-氮-2′-脱氧胞苷处理前后A375和M14细胞中IGFBP7mRNA和蛋白的表达，噻唑蓝法检测不同浓度5-氮-2′-脱氧胞苷对A375和M14细胞的生长抑制作用。结果 10 μmol/L 5-氮-2′-脱氧胞苷处理A375和M14细胞48 h后，IGFBP7 mRNA和蛋白水平的表达均得到恢复。不同浓度的5-氮-2'-脱氧胞苷分别作用24、48、72、96和120 h后，呈时间（F = 141.35，P < 0.01；F = 549.33，P < 0.01）和剂量（F = 561.12，P < 0.01；F = 271.43，P < 0.01）依赖性抑制A375和M14细胞增殖。结论 DNA甲基化参与调控IGFBP7基因的表达，5-氮-2′-脱氧胞苷具有抑制黑素瘤细胞增殖的作用。

关键词: 5-氮杂-2-脱氧胞苷酸, 胰岛素样生长因子结合蛋白7, 黑素瘤

Abstract:

Objective To study the effects of 5-aza-dc on the expression of IGFBP7 in and proliferation of melanoma cell lines A375 and M14. Methods Reverse transcription-PCR and immunocytochemistry were performed to detect the mRNA and protein expression of IGFBP7 in A375 cells and M14 cells after treatment with 5-aza-dc of 10 μmol/L for 48 hours, and MTT assay to measure the proliferation of both cell lines treated with 4 different concentrations （2.5, 5, 10, 20 μmol/L） of 5-aza-dc for various durations. Results The treatment with 5-aza-dc restored IGFBP7 expression at both mRNA and protein levels. The four concentrations of 5-aza-dc inhibited the proliferation of A375 and M14 cells in a dose-dependent （F = 561.12, 271.43, respectively, both P < 0.01) and time-dependent （F = 141.35, 549.33, respectively, both P < 0.01） manner. Conclusions DNA methylation may be involved in the modulation of aberrant IGFBP7 gene expression in melanoma, and 5-aza-dc could inhibit the proliferation of A375 and M14 cells.

Key words: 5-aza-dc, IGFBP7, melanoma