摘要
目的:研究人参单体皂苷Rh2对激素非依赖性人前列腺癌细胞(PC-3M细胞株)的致凋亡作用及其对新基因GRIM-19表达的影响。方法:实验分为Rh2高剂量组(40 mg.L-1)、Rh2中剂量组(30 mg.L-1)、Rh2低剂量组(15 mg.L-1)及对照组(未加药组)。分别作用于PC-3M细胞24 h,吖啶橙染色观察其对PC-3M细胞的促凋亡作用;RT-PCR和Western blotting方法分别检测Rh2对PC-3M细胞GRIM-19 mRNA及其蛋白表达的影响。结果:Rh2高、中、低剂量组作用于PC-3M细胞24h吖啶橙染色均呈阳性结果。Rh2低剂量组出现典型“出芽”现象,细胞中出现致密鲜亮橙色斑点。对照组细胞呈均匀绿色;RT-PCR结果显示,GRIM-19 mRNA随Rh2浓度的增加表达逐渐增强,Rh2中、低剂量组与对照组比较差异具有显著性(P<0.05),Rh2高剂量组GRIM-19 mRNA表达与对照组比较差异具有高度显著性(P<0.01)。Western blotting结果与RT-PCR结果相一致,GRIM-19蛋白随Rh2浓度的增加其表达逐渐增强,Rh2中低剂量组与对照组比较差异具有显著性(P<0.05),Rh2高剂量组GRIM-19蛋白的表达与对照组比较差异具有高度显著性(P<0.01)。结论:Rh2诱导PC-3M细胞凋亡与上调GRIM-19基因和蛋白表达有关,其表达强度呈剂量依赖性。
Objective To study the effects of Rh2 （a ginsenoside extracted from ginseng） on PC-3M （androgen independent cell lines） apopotosis and new gene GRIM-19 expression. Methods Rh2 agents were divided into three groups： high dose group （Rh240 mg · L^-1）, middle dose group （Rh2 30 mg · L^-1） and low dose group （Rh2 15 mg · L^-1）,which incubated with PC-3M for 24 h, respectively, in the following experiments. Morphological assessment of apoptosis was performed by acridine orange （AO） stained fluorescence microscope to detect the apoptosis of PC-3M cells after treated with Rh2 agents; RT-PCR was used to measure the level of GRIM-19 mRNA in PC-3M cells treated with Rh2 agents; GRIM-19 protein expression of PC-3M cells treated with Rh2 agent was measured by Western blotting. Results After various doses of Rh2 interacted with PC-3M cells for 24 h, AO assay showed that cells changed from green color into orange one and typical exocytosis phenomena were observed in Rh2 15 mg · L^-1 group. The consequences of RT-PCR and Western blotting demonstrated that GRIM-19 expression was intensified both in gene and protein levels with the increasing Rh2 doses. The expressions of GRIM-19 in Rh2 15 mg · L^-1 group and 30 mg · L^-1 group were higher than that in control group （P〈0.05）, and in Rh2 40 mg · L^-1 group the expression of GRIM-19 had a remarkable difference compared with control group （P〈 0.01）. Conclusion Rh2 can lead to apoptosis of PC-3M cells that correlated with GRIM-19 gene and protein expression upregulation, the expression level increases with the Rh2 in a dose-dependent manner.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2006年第2期186-188,193,共4页
Journal of Jilin University：Medicine Edition
基金
国家科技部国际科技合作重点项目(2004DFB02000)
中日政府间专项技术合作项目(第59号)
