【摘要】 目的:探讨茉莉酸甲酯(MJ)对胃癌SGC7901细胞生长的抑制作用及其机制。方法:四甲基偶氮唑(MTT)法检测细胞生长活性;Hoechst 33258荧光染色法观察凋亡细胞核的形态学变化;AnnexinⅤ-FITC/PI双染流式细胞术检测细胞的凋亡率;蛋白质印迹法检测凋亡蛋白Caspase-3和Caspase-9的表达。结果:MTT法显示,MJ对胃癌SGC7901细胞生长有显著抑制作用,且呈剂量-时间依赖关系,0.5、1.0和2.0mmol/L MJ作用6h后,抑制率分别为(7.10±0.84)%、(18.22±1.82)%和(26.89±1.56)%,溶剂对照组为(0.43±0.26)%,顺铂(DDP)组为(10.44±1.17)%,总体比较差异有统计学意义(F=232,P=0.000),各组间比较差异均有统计学意义,P值均<0.01;采用MJ上述3种浓度作用12h后,抑制率分别为(12.44±1.20)%、(32.70±1.29)%和(52.94±2.29)%,溶剂对照组为(0.31±0.41)%,DDP组为(22.60±1.97)%,总体比较差异有统计学意义(F=1 013,P=0.000),各组间比较差异均有统计学意义,P值均<0.01;MJ三种浓度作用24h后,抑制率分别为(17.37±1.95)%、(41.79±4.40)%和(71.93±4.65)%,溶剂对照组为(0.34±0.21)%,DDP组为(27.72±2.45)%,总体比较差异有统计学意义(F=444,P=0.000),各组间比较差异均有统计学意义,P值均<0.01;其中以浓度为2.0mmol/L MJ作用24h后,对SGC7901细胞的抑制率效果最佳。荧光染色法观察MJ能够诱导人胃癌SGC7901细胞株的凋亡。流式细胞检测显示,MJ可促进胃癌SGC7901细胞的凋亡,凋亡率为(68.14±7.91)%;与溶剂对照组(6.98±1.45)%相比,差异有统计学意义,P=0.013。蛋白质印迹法检测显示,MJ可上调凋亡蛋白Caspase-3和Caspase-9的表达;事先采用辣椒素受体拮抗剂(Cap)预处理,MJ的抗胃癌SGC7901细胞的效应可被消除,与MJ单独给药组相比,MJ对癌细胞的生长抑制率下降为(38.79±2.77)%,P=0.05;凋亡细胞明显减少;肿瘤细胞凋亡率降为(39.81±4.57)%,P=0.045;凋亡蛋白Caspase-3和Caspase-9表达也降低。结论:MJ有显著抑制胃癌SGC7901细胞增殖的作用;辣椒素受体(VR1)可能介导了MJ对SGC7901细胞增殖的抑制作用,使细胞内凋亡信号通路激活,促进了肿瘤细胞的凋亡。更多还原

【Abstract】 OBJECTIVE: To explore the growth inhibitory effects of methyl jasmonate(MJ) on human gastric cancer SGC7901 cell line,and investigate its mechanisms.METHODS: The MTT colorimetry was used to detect the growth inhibitory rates of MJ on human gastric cancer SGC7901 cell;Hoechst 33258 fluorochrome staining was employed to observe cell apoptosis;the apoptosis potential of cells was assessed by flow cytometry with Annexin Ⅴ-FITC/PI.The expressions Caspase-3 and Caspase-9 were detected by Western blott.RESULTS:MJ inhibited the growth of SGC7901 cells in a dose-and time-dependent manner.After exposed to 0.5,1.0 and 2.0 mmol/L MJ for 6 h,the inhibition rates were about(7.10±0.84)%,(18.22±1.82)% and(26.89±1.56)% respectively,compared with the solvent control group(0.43±0.26)% and DDP group(10.44±1.17)%,there was a statistically significant difference on total(F=232,P=0.000) and each group(P<0.05);With the same dose MJ for 12h,the inhibition rates were about(12.44±1.20) %,(32.70± 1.29) %and(52.94±2.29) % respectively.Compared with the solvent control group(0.31±0.41) % and DDP group(22.60±1.97) %,there was a statistically significant difference on total(F=1 013,P=0.000) and each group(P< 0.01);for 24h,the inhibition rates were about(17.37±1.95) %,(41.79±4.40) % and(71.93±4.65) %,respectively.Compared with the solvent control group(0.34±0.21) % and DDP group(27.72±2.45) %,there was a statistically significant difference on total(F=444,P=0.000) and each group(P<0.01).2mol / L MJ after 24hhad the best inhibition rates.Hoechst 33258fluorochrome staining showed that there were many apoptotic cells in MJ group.The apoptosis rate was(68.14±7.91) %,after exposed to 2.0mmol / L MJ for 24h,compared with the solvent control group(6.98±1.45) %(P=0.013).The expressions of Caspase-3and Caspase-9protein were increased.Above-mentioned inhibitory effects of MJ on SGC7901cell were abolished,respectively by the pretreatment with capsazepine,an antagonist of vanilloid receptor subtype 1.The percentage of inhibition rate and apoptosis dropped to(38.79±2.77) %and(39.81±4.57) %(P=0.05;P=0.045),respectively.The expressions of protein were also decreased.CONCLUSIONS : MJ can significantly inhibit the growth of SGC7901cells through inducing cell apoptosis.Its molecular mechanism was probably associated with vaniloid receptor subtype 1(VR1) signal pathway.更多还原