摘要
目的探讨人参总皂甙（total saponins of panax ginseng,TSPG）对人急性髓系白血病细胞株KG1α增殖的影响。方法取对数生长期的KG1α细胞,调整密度为5×108/L,空白对照组予以常规培养;人参总皂甙TSPG组分别加入0.1、0.2、0.4、0.6、0.8 g/L TSPG。采用MTT比色法测定细胞存活率,瑞氏染色法以及光电镜观察细胞形态学改变。流式细胞仪测定细胞周期分布的变化以及细胞的凋亡率。免疫细胞化学的方法检测细胞中β-catenin、NF-κB p65蛋白表达的变化。RT-PCR半定量检测细胞中β-catenin mRNA和NF-κB p65 mRNA水平的变化。结果人参总皂甙TSPG在体外对KG1α增殖具有明显抑制作用。在0.1~0.8 g/L TSPG浓度范围内,其抑制作用呈浓度依赖性,在浓度0.4 g/L处达到细胞的半数抑制率,且在48 h抑制率达到高峰。0.4 g/L TSPG作用48 h后,可见细胞分散生长,数量明显减少;细胞明显分化或凋亡,产生核分裂象,胞质中产生凋亡小体以及空泡;与对照组相比,加药组KG1α细胞周期分布无明显变化（P〉0.05）;与对照组相比,加药组细胞凋亡率显著增高,其改变具有统计学意义（P〈0.05）;与对照组相比,加药组细胞内β-catenin蛋白因子表达降低其表达区域由胞核向胞膜转移,NF-κB p65因子的表达也明显减弱其表达区域由胞核向胞膜转移;与对照组相比,加药组β-catenin和NF-κB p65 mRNA的水平降低,其改变具有统计学意义（P〈0.05）。结论 TSPG可抑制KG1α细胞增殖诱导细胞凋亡,且TSPG抑制KG1α细胞增殖的作用机制可能与其抑制β-catenin及NF-κB p65蛋白表达,降低β-catenin和NF-κB p65 mRNA的水平有关。
Objective To investigate the effect of total saponins of panax ginseng（TSPG） on the proliferation of acute myelocytic leukemia KG1α cells.Methods The density of KG1α cells with logarithmic phase were regulated to 5×108/L and incubated with 0.1,0.2,0.4,0.6,and 0.8 g/L TSPG respectively.The conventional culture was carried out in the blank control group.The effects of TSPG on the proliferation of KG1a cells were examined by MTT assay.Morphological alterations of KG1a cells were observed by light microscopy and electron microscopy.The effect of TSPG on KG1a cell cycle and apoptosis were determined by flow cytometry（FCM）.Expression of β-catenin and NF-κB p65 protein was detected by immunohistochemistry.Expressions of β-catenin and NF-κB p65 mRNA were detected by RT-PCR.Results TSPG inhibited KG1α cells proliferation significantly in vitro in a dose-dependent manner at range of 0.1 to 0.8 g/L.And the efficiency reached its peak at the dose of 0.4 g/L for 48 h.Compared with the blank control,apoptotic cells and morphologic changes were found under microscope after the cell were treated with 0.4 g/L TSPG for 48 h.The analysis of cell cycle showed that there was no significant change compared with that of the blank control（P0.05）.The apoptotic rate of cells was significantly raised compared with that of the blank control（P0.05）.The expressions of β-catenin and NF-κB p65 proteins were decreased in KG1α cells.The levels of mRNA of β-catenin and NF-κB p65 were declined compared with the blank control（P0.05）.Conclusion TSPG inhibits the proliferation of KG1α cells by inducing apoptosis,probably by the mechanism of inhibiting the expressions of β-catenin and NF-κB at protein and mRNA levels.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第24期2599-2603,共5页
Journal of Third Military Medical University
基金
国家自然科学基金(30873406)~~
关键词
人参总皂甙
KG1α细胞
增殖抑制
凋亡
total saponins of panax ginseng
KG1α cells
inhibit the proliferation
apoptosis
