8-硝基白杨素诱导HL-60细胞凋亡和抑制增殖与PTEN-Akt途径的相关性研究  

Relationship between PTEN-Akt pathway and 8-nitrochrysin induced apoptosis and inhibition of the proliferation of HL-60 cells

作　　者：岳军[1] 

机构地区：[1]邵阳医学高等专科学校生物化学教研室,湖南邵阳422000

出　　处：《华西药学杂志》2009年第2期142-144,共3页West China Journal of Pharmaceutical Sciences

基　　金：湖南省教育厅优秀青年项目资助(06B087)

摘　　要：目的观察8-硝基白杨素(NOChR)抑制人急性髓性白血病细胞系(HL-60)细胞增殖和诱导凋亡的作用,并探讨其作用机制。方法体外培养HL-60细胞;MTT比色法检测增殖活性;流式细胞术定量分析细胞凋亡程度;琼脂糖凝胶电泳观察DNA梯形条带。Western Blot分析HL-60细胞PTEN、P-Akt蛋白表达的影响。结果NOChR可显著抑制HL-60细胞增殖,呈浓度依赖性,IC50为1.34μmol;NOChR具有诱导HL-60细胞凋亡的作用,且呈浓度和时间依赖性。NOChR以浓度和时间依赖方式上调HL-60细胞PTEN蛋白的表达和抑制P-Akt蛋白的表达,其作用可被PPARγ特异性阻断剂GW9662(10μmol)预孵育拮抗。结论NOChR诱导HL-60细胞凋亡作用与其活化PPARγ、上调PTEN蛋白表达、抑制Akt磷酸化相关。OBJECTIVE To observe the inhibition effect of 8 - nitrochrysin on the proliferation of HL - 60 cell and induction of HL -60 cell apoptosis, and investigate the action mechnism. METHODS HL -60 cell line was cultured in vitro and the proliferative activity of HL - 60 cells treated with 5,7 - dihydrox - 8 - nitrochrysin was measured using 3 - （4,5 - dimethyhhiazol - 2 - yl） - 2,5 - dipbenytertrazolium blue （MTT） colorimetric assay. The apoptosis rate of HL - 60 cell line induced by NOChR was analyzed using flow cytometry （FCM） with PI staining. DNA ladder bands were observed by DNA agarose gel electrophoresis. The influence of NOChR on the PTEN and p - AKt protein expression of HL - 60 cells was analyzed with western blot. RESULTS The proliferation of HL -60 cells was markedly inhibital by NOChR in a concentration -dependent manner, and the IC50 was 1.34 μ mol. Flow cytometry after PI staining indicated that apoptosis in HL -60 cells was induced by NOChR in a time and concentration dependent manner. Western blot indicated that NOChR can efficiently up -regulated PTEN and down -regulated p- AKt protein expression of HL- 60 cells in a concentration and time dependent manner, and the pre - incubation of GW9662, blocker of PPARγ, can antagonize the up - regulating effect of 8 - nitrochrysin on PTEN protein expression of HL - 60 cells. CONCLUSION NOChR induced apoptosis of HL - 60 cells may be associated with activation of PPARγ, up - regulation of PTEN and inhibition of p - AKt by 8 - nitrochyrsin.

关 键 词：人急性髓性白血病 白杨素 凋亡 细胞增殖 PTEN蛋白表达 HL-60细胞 

分 类 号：R96[医药卫生—药理学]