刘艺,李静,刘晓岩,姜蓉,王建伟,魏强,赵亮,陈地龙
LIU Yi,LI Jing,LIU Xiao-yan,JIANG Rong,WANG Jian-wei,WEI Qiang,ZHAO Liang,CHEN Di-long(Laboratory of Stem Cell and Tissue Engineering

• 1:重庆医科大学干细胞与组织工程研究室组织学与胚胎学教研室

摘要(Abstract)：

目的探讨人参皂苷单体Rh2(S亚型)对人急性髓系白血病细胞株KG1α增殖、凋亡的影响及其机制。方法取对数生长期的KG1α细胞,分为两组:空白对照组(常规培养)和人参皂苷单体Rh2(S)组,采用MTT法检测细胞的增殖活力,倒置显微镜观察细胞的形态及数量,透射电镜观察细胞的超微结构,流式细胞仪检测细胞周期的分布,Real-time PCR检测细胞中β-catenin基因mRNA水平,免疫细胞化学法检测细胞中β-catenin、TCF4、CyclinD1蛋白的定位及表达,Western blot检测细胞中β-catenin、TCF4、CyclinD1蛋白的表达水平。结果人参皂苷单体Rh2(S)对KG1α细胞的增殖具有明显的抑制作用,且呈浓度和时间依赖性。与空白对照组比较,Rh2(S)作用48 h后,可见KG1α细胞分散生长,数量明显减少;可见数量不等,程度不同的凋亡细胞;G0/G1期细胞比例明显上升(P<0.05),G2+M和S期细胞比例明显下降(P<0.05);细胞中β-catenin基因mRNA水平明显降低(P<0.01)。β-catenin、TCF4、CyclinD1蛋白的表达水平明显降低(P<0.05或P<0.01)。结论人参皂苷单体Rh2(S亚型)对KG1α细胞具有明显的增殖抑制作用,其机制可能抑制Wnt/β-catenin/TCF4/CyclinD1信号通路,使细胞周期阻滞,诱导细胞凋亡。
Objective To investigate the effect of ginsenoside Rh2(subtype S)on proliferation and apoptosis of acute myelocytic leukemia KG1α cell strain as well as the relevant mechanism.Methods KG1α cells at logarithmic phase were divided into two groups.The cells in blank control group were subjected to routine culture,while those in test group were incubated with Rh2(S)at various concentrations.The cells were determined for proliferative activity by MTT method,observed for morphology and count under invert microscope and for ultrastructure under transmission electron microscope,and determined for distribution of cell cycles by flow cytometry,for β-catenin mRNA level by Real-time PCR,for location and expression of β-catenin,TCF4,and cyclinD1 by immunocytochemical method,and for expression levels of the proteins by Western blot.Results Rh2(S)inhibited the proliferation of KG1α cells significantly,in dose-and time-dependent manners.Compared with those in blank control,the KG1α cells 48 h after treatment with 0.04 g/L Rh2(S) grew in scatter,of which the count decreased significantly.Cells at different counts and apoptosis levels were observed.The percentage of cells in G0/G1 phase increased significantly(P < 0.05),while those in G2 + M and S phases decreased significantly(P < 0.05).The β-catenin mRNA level(P < 0.01)and the expression levels of β-catenin,TCF4 and cyclinD1 protein decreased significantly(P < 0.05 or P < 0.01).Conclusion Ginsenoside Rh2(S)inhibited the proliferation of KG1α cells significantly by a possible mechanism of inhibiting Wnt/β-catenin/TCF4/CyclinD1 signaling pathway thus causing cell cycle arrest and inducing cell apoptosis.

关键词(KeyWords)： 人参皂苷单体Rh2(S亚型);KG1a细胞;细胞增殖;细胞凋亡;β-catenin;TCF4;CyclinD1
Ginsenoside Rh2(subtype S);KG1α cells;Cell proliferation;Cell apoptosis;β-catenin;TCF4;CyclinD1

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(81171929);; 重庆市自然科学基金项目(CSCT 2009BB5075);; 重庆市教委资助课题(Kj080302)

作者(Author): 刘艺,李静,刘晓岩,姜蓉,王建伟,魏强,赵亮,陈地龙
LIU Yi,LI Jing,LIU Xiao-yan,JIANG Rong,WANG Jian-wei,WEI Qiang,ZHAO Liang,CHEN Di-long(Laboratory of Stem Cell and Tissue Engineering

DOI: 10.13200/j.cjb.2012.10.80.liuy.020

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