蒿甲醚对人胶质瘤细胞系U251细胞凋亡的影响

Effect of artemether on cell cycle and apoptosis of human glioblastoma cell line U251 cells

目的研究蒿甲醚对体外培养的人胶质瘤细胞系U251细胞凋亡的影响并初步探讨其机制。方法不同浓度蒿甲醚(25、50、100、200、400、800 μmol/L)作用U251胶质瘤细胞,四甲基偶氮唑盐法测定药物抑制率和半抑制浓度(IC50);流式细胞术检测细胞周期的变化和细胞凋亡情况;Hoechst33258荧光染色检测细胞凋亡情况。结果蒿甲醚对U251细胞生长抑制率随药物浓度增加而显著增加(P<0.05),同时随药物作用时间延长而显著增加(P<0.05)。药物作用24、48和72 h,其IC50分别为849.28±25.89、518.93±32.83和172.99±6.06μmol/L,三者之间均差异显著(P<0.05)。经400 μmol/L蒿甲醚作用48 h,Hochest33258荧光染色可观察到凋亡小体。经400 μmol/L蒿甲醚作用24、48、72 h,细胞凋亡率分别为(4.55±0.24)%、(12.82±1.88)%和(24.22±2.17)%,三者之间均差异显著(P<0.05);细胞周期分析显示,随着蒿甲醚作用时间的延长,G0/G1期细胞比例显著增加(P<0.05),S期和G2/M期细胞比例显著减少(P<0.05)。结论蒿甲醚能抑制U251细胞生长,呈剂量和时间依赖性;其机制可能为将细胞阻滞在G0/G1期并诱导其凋亡。

Objective To investigate the effect of artemether on cell cycle and apoptosis of human gliblastoma cell line U251 cells. Methods The U251 cells were cultured respectively in the media containing artemether of different concentrations （25, 50, 100, 200, 400, 800 μmol/L） for 24, 48 and 72 h. The inhibition rate （IR） of U251 cells growth produced by artemether was determined by methylthiazolyl tetrazoliurn test, and the proportion of U251 cells in the different phases of cell cycle was analyzed by the flow cytometry （FCM）. The apoptosis of U251 cells was analyzed by Hochest33258 fluorescent staining and FCM. Results The IR significantly increased when the concentration of artemether or the cultural time increased （P〈0.05）. The IRs were （34.72±2.33）%, （45.08±2.26）% and （58.00±2.95）% respectively when the U251 cells were treated with artemether at concentration of 400 μmol/L for 24, 48 and 72 h. The Hoechst33258 staining showed that the apoptotic bodies were seen in the U251 cells treated with artemether at the concentration of 400 μmol/L for 48 h. After the treatment with artemether at the concentration of 400 μmol/L for 24, 48 and 72 h, the U251 cell apoptosis rates were （4.55±0.24）%, （12.82± 1.88）% and （24.22±2.17）% respectively, whereas the proportion of G0/G1 cells significantly increased （P〈0.05） and the proportion of S and G2/M cells significantly decreased （P〈0.05） when the cultural time increased. Conclusions That artemether can inhibit the proliferation of U251 cells in a dose- and time-dependent manner may be produced by blocking cells at the G0/G1 phase and inducing cell apoptosis.