吉林大学学报(医学版) ›› 2006, Vol. 32 ›› Issue (2): 186-188.doi: 科技部国际科技合作重点项目(2004DFB02

• 基础研究 • 上一篇    下一篇

人参皂苷Rh2诱导PC-3M细胞凋亡及其对新基因GRIM-19表达的影响

邵月婷1,李 峰1,高丽芳1,孙连坤1,胡嘉弟2,李 扬1*,赵雪俭1   

  1. 1. 吉林大学基础医学院病理生理学教研室 吉林大学前列腺疾病防治研究中心,吉林 长春130021; 2. 美国马里兰大学医学院,美国21201
  • 收稿日期:2005-08-17 修回日期:1900-01-01 出版日期:2006-03-28 发布日期:2006-03-28
  • 通讯作者: 李 扬1*

Apoptosis of PC-3M cells induced by Rh2 and effectsof Rh2 on expression of GRIM-19

SHAO Yue-ting1,LI Feng1, GAO Li-fang1, SUN Lian-kun1,HU Jia-di2, LI Yang1*,ZHAO Xue-jian1   

  1. 1. Department of Pathophysiology of Basic Medical Sciences,Prostate Diseases Prevention and Treatment Research Center, Jilin University, Changchun 130021,China; 2. School of Medical Sciences, University of Maryland, MD 21201, USA
  • Received:2005-08-17 Revised:1900-01-01 Online:2006-03-28 Published:2006-03-28
  • Contact: LI Yang1*

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。

Abstract: 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 (Rh230 mg·L-1)and low dose group(Rh215 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.

中图分类号: 

  • R363