雷公藤内酯醇对人黑素瘤M14细胞系增殖与凋亡的影响

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

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、30nmol／L3种浓度雷公藤内酯醇作用48h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30nmoYL雷公藤内酯醇作用48h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、30nmol/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30nmol/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。

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.