三七总皂苷抑制K562细胞mTOR信号通路活性的实验研究  

Panax notoginseng saponins suppresses m TOR signaling pathway in K562 cells

作　　者：翟玮玮[1] 李玉云[2] 杨向荣[1] 于北凯 王露[1] 李见[1] 

机构地区：[1]蚌埠医学院临床检验诊断学实验中心,安徽蚌埠233030 [2]蚌埠医学院临床检验基础教研室,安徽蚌埠233030

出　　处：《第三军医大学学报》2015年第12期1237-1242,共6页Journal of Third Military Medical University

基　　金：安徽省教育厅自然科学基金(KJ2012A198)~~

摘　　要：目的探讨三七总皂苷（Panax notoginseng saponins,PNS）对体外培养K562细胞m TOR信号通路相关分子表达及活性的影响。方法不同浓度的PNS（50~800μg/m L）干预体外培养K562细胞24~72 h后,MTT比色法检测PNS对K562细胞增殖的抑制作用;AO/EB双荧光染色法观察细胞死亡及凋亡状况;RT-PCR检测m TOR、p70S6K、4E-BP1 mRNA表达变化;Western blot检测m TOR、p70S6K、4E-BP1及p-m TOR、p-p70S6K、p-4E-BP1蛋白表达量的变化。结果与对照组相比,浓度为100~800μg/m L的PNS能够抑制K562细胞增殖（P〈0.05）,促进K562细胞凋亡及死亡（P〈0.05）,PNS对K562细胞72 h的IC50为（229.07±2.36）μg/m L;100、200、400μg/m L PNS作用于K562细胞72 h后m TOR蛋白表达量及mRNA表达量降低（P〈0.05）,且磷酸化蛋白p-m TOR（Ser2448）、p-p70S6K（Thr229/389）及p-4E-BP1（Thr37/46）表达水平也随着药物浓度的增加而降低（P〈0.05）。结论 PNS能够抑制K562细胞m TOR信号通路的活性,这可能是PNS抑制K562细胞增殖的机制之一。Objective To investigate the effect of Panax notoginseng saponins（ PNS） on the expression and activity of m TOR signaling pathway related molecules in K562 cells in vitro. Methods K562 cells were treated with different concentrations（ 50- 800 μg / m L） of PNS for different periods of time（ 24-72 h）. The cell proliferation was measured by MTT assay,and the cell death and apoptosis were observed by AO / EB double fluorescence staining. The mRNA expression of m TOR,p70S6 K and 4E-BP1 was examined by RT-PCR,and the protein expression of m TOR, p70S6 K, 4E-BP1 and their phosphorylated proteins（ p-m TOR,p-p70S6 K and p-4E-BP1） was detected by Western blotting. Results PNS（ 100- 800 μg/m L）inhibited the proliferation of K562 cells（ P 〈 0. 05） and promoted cell apoptosis and death（ P 〈 0. 05）,and72-h IC50was（ 229. 07 ± 2. 36） μg / m L. After PNS（ 100,200 and 400 μg / m L） was applied to K562 cells for 72 h,the mRNA and protein expressions levls of m TOR were dose-dependently reduced（ P 〈 0. 05）. In addition,the expression of p-m TOR（ Ser2448）,p-p70S6K（ Thr229 /389） and p-4E-BP1（ Thr37 /46）proteins was also decreased in a dose-dependent manner（ P 〈 0. 05）. Conclusion PNS can inhibit the activity of m TOR signaling pathway in K562 cells,and this may be one of PNS-mediated cell proliferation inhibition mechanisms.

关 键 词：三七总皂苷 K562细胞 M TOR信号通路 

分 类 号：R285.5[医药卫生—中药学] R73-354[医药卫生—中医学]