中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (9): 641-643.

• 论著 • 上一篇    下一篇

雷公藤内酯醇对人黑素瘤M14细胞系增殖与凋亡的影响

陶玥1,马鹏程2,孙建方2,周武庆1,3   

  1. 1. 中国医学科学院北京协和医学院皮肤病研究所
    2. 南京 中国医学科学院北京协和医学院皮肤病研究所
    3.
  • 收稿日期:2012-04-01 修回日期:2012-06-11 出版日期:2012-09-15 发布日期:2012-08-31
  • 通讯作者: 马鹏程 E-mail:mpc815@163.com

Effects of triptolide on the proliferation of and apoptosis in a human melanoma cell line M14

  • Received:2012-04-01 Revised:2012-06-11 Online:2012-09-15 Published:2012-08-31
  • Contact: MA Peng-Cheng E-mail:mpc815@163.com

摘要:

目的 探讨雷公藤内酯醇对体外培养人黑素瘤M14细胞增殖与凋亡的影响。 方法 利用CCK8检测不同浓度和时间雷公藤内酯醇作用下,对黑素瘤M14细胞的增殖抑制作用;用10、20、30 nmol/L 3种浓度雷公藤内酯醇作用48 h后,以流式细胞仪检测细胞周期变化、Annexin-Ⅴ/PI法观察M14细胞凋亡的变化;用Hoechst33258染色法观察30 nmol/L雷公藤内酯醇作用48 h后细胞凋亡的形态变化。结果 细胞周期显示,雷公藤内酯醇处理过的M14细胞S期比例高于对照组, G2/M期比例低于对照组。空白对照组及10、20、30 nmol/L浓度雷公藤内酯醇处理的M14细胞凋亡率分别为(2.92 ± 0.17)%,(20.99 ± 0.40)%, (34.28 ± 2.04)% 和(63.38 ± 0.71)%,呈剂量依赖性诱导M14细胞凋亡,经30 nmol/L雷公藤内酯醇作用后,Hoechst33258染色示凋亡细胞形态改变。 结论 雷公藤内酯醇对黑素瘤M14细胞具有抑制增殖和诱导凋亡的作用。

关键词: 凋亡

Abstract:

Objective To study the effects of triptolide on the apoptosis in and proliferation of a human melanoma cell line M14. Methods M14 cells were cultured with the presence of 5 concentrations (12.5, 25, 50, 100, 200 nmol/L) of triptolide for 24, 48 and 72 hours respectively, and cell counting kit-8 (CCK-8) was used for the detection of cell proliferation. Some M14 cells were treated with triptolide at 10 nmol/L, 20 nmol/L and 30 nmol/L for 48 hours followed by the analysis of cell cycle by flow cytometry and detection of cell apoptosis by flow cytometry following annexin V- fluorescein isothiocyanate (FITC)/propidium iodide double staining. The morphological changes of M14 cells treated by triptolide at 30 nmol/L for 48 hours were observed by Hoechest 33258 staining. Results Compared with untreated M14 cells, an increase of cell population in S phase was observed in triptolide-treated cells,along with a decline in cell population in G2/M phase. The apoptosis rate was (2.92 ± 0.17)%, (20.99 ± 0.40)%, (34.28 ± 2.04)% and (63.38 ± 0.71) % respectively in M14 cells treated with triptolide at 0, 10, 20 and 30 nmol/L for 48 hours, suggesting that triptolide enhanced the proliferation of M14 cells in a dose-dependent manner. After treatment with triptolide of 30 nmol/L, M14 cells showed morphological changes characteristic of apoptosis. Conclusion Triptolide could inhibit the proliferation of and induce the apoptosis in M14 human melanoma cells.

Key words: Apoptosis