土槿皮乙酸诱导大鼠神经胶质瘤C6细胞株凋亡  

Study on action mechanisms for the effect of pseudolaric acid B in inducing apoptosis of murine C6 glioma cell line

作　　者：高芹 吴侠[2] 赵运胜[2] 

机构地区：[1]秦皇岛市抚宁区人民医院检验科,河北秦皇岛066300 [2]秦皇岛市第一医院检验科,河北秦皇岛066000

出　　处：《现代肿瘤医学》2017年第11期1712-1716,共5页Journal of Modern Oncology

摘　　要：目的:研究土槿皮乙酸诱导大鼠神经胶质瘤C6细胞株凋亡及其机制。方法:采用MTT法检测0、0.625、1.25、2.5、5、10、20、40、80μmol/L不同终浓度PAB对C6细胞增殖能力的影响,激光扫描共聚焦显微镜观察AO/EB荧光染色后细胞的凋亡情况,流式细胞术检测碘化丙啶(PI)染色后细胞周期的变化,免疫组化法测定C6细胞的凋亡相关蛋白Bcl-2的表达。结果:MTT结果显示,PAB呈时间剂量依赖性抑制C6细胞的生长。PAB处理24、48和72 h后,对C6细胞的半效致死剂量(IC50)分别为39.811、23.306和7.360μmol/L。5.0、10.0、20.0μmol/L PAB作用C6细胞72 h后的凋亡率分别为(50.5±0.7)%、(63.5±2.3)%和(80.0±1.2)%,与对照组(12.0±1.6)%比较均明显升高,差异有统计学意义(P<0.05)。流式细胞仪检测结果显示5.0、10.0、20.0μmol/L不同浓度PAB作用C6细胞72小时后,随着浓度的增加,G2/M期细胞逐渐增多,分别为(29.84±6.215)%,(34.58±1.187)%和(70.23±6.325)%,与对照组(7.79±0.570)%相比,后两组差异有统计学意义(P<0.05)。C6细胞中Bcl-2蛋白阳性表达率随药物剂量的增大而减小。结论:PAB可抑制大鼠神经胶质瘤C6细胞增殖,阻滞细胞周期于G2/M期、下调Bcl-2表达为其可能的作用机制。Objective： To investigate the action mechanisms for the effect of pseudolaric acid B in inducing apopto-sis of murine C6 glioma cell line. Methods ： The effects of different mass concentrations of PAB （ 0 ,0 . 6 2 5 ,1 .2 5 ,2 . 5 , 5,10,20,40,80 μmol/L） on the growth of C6 cells were analysed by MTT assay. The apoptosis effect of C6 cells was detected by inverted UV fluoresence microscope with AO/EB staining. Cell cycle was checked by flow cytometry with PI staining. The expression of Bel -2 protein （apoptosis associated protein） was determined by immunohisto - chem-istry. Results： MTT assay revealed PAB inhibited the growth of C6 cells in a dose and time dependent manner. The median lethal dosages （IC50） of PAB on C6 cells were 39. 811,23. 306 and 7. 360 jjimol/L after treatment with PAB for 24,48 and 72 h respectively. The apoptosis rates were （50. 5 ±0. 7）%,（ 63.5 ±2.3）% and （80. 0 ± 1.2） % af-ter treatment with PAB （5.0,10. 0,20. 0 jjimol/L） for 72 h respectively, they were all significantly different from that of control group （12. 0 ± 1.6） % （P 〈 0 . 0 5 ） . Flow cytometry indicated that the increasing of C6 cells in G2/M phase occurred 72 h after treatment with increasing of the concentration of PAB （5.0,10. 0 and 20. 0 jjimol/L） , and the rates were （29. 84 ±6.215）%,（ 34. 58 ±1. 187）% and （70. 23 ± 6. 325 ） % respectively,the last two were signifi-cantly different from that of control group （7. 79 ±0. 570）% （P 〈0. 05）. The expression of Bel -2 protein in C6cells decreased with the increasing of concentration of PAB. Conclusion ： PAB can inhibit the growth of murine glioma C6 cells and induce cell apoptosis, which may be associated with the decreased expression of Bel -2 and arrested in G2/M phase.

关 键 词：土槿皮乙酸 神经胶质瘤C6细胞 细胞凋亡 

分 类 号：R730.264[医药卫生—肿瘤] R739.41[医药卫生—临床医学]