NF-κB抑制剂PDTC抗白血病细胞增殖的实验研究  

Effect of NF-κB inhibitor pyrrolidine dithiocarbonate on the proliferation and apoptosis in K562 cells

作　　者：张纯[1] 崔国惠[1] 刘芳[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,湖北武汉430022

出　　处：《中国病理生理杂志》2009年第12期2349-2352,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30700882)

摘　　要：目的:观察核转录因子NF-κB活性特异性抑制剂PDTC(吡咯烷二硫代氨基甲酸盐)对急性白血病细胞系K562细胞增殖的影响,并探讨其机制。方法:利用Trans AMTM NF-κB/p65活性测定试剂盒,检测PDTC作用K562细胞12h、24h后NF-κB亚基p65活性的变化;采用WST-1细胞增殖试验观察不同浓度PDTC作用24h、48h、72h对细胞增殖的影响;用单细胞凝胶电泳(彗星检测)方法检测PDTC对K562细胞DNA损伤的影响;利用免疫印迹法(Western blotting)检测PDTC对K562细胞中pro-caspase-3和活化型caspase-3蛋白表达的影响。结果:与对照组相比,经PDTC处理的实验组K562细胞NF-κB/P65的活性受到明显抑制(P<0.01);且PDTC能以时间和剂量依赖方式抑制K562细胞的增殖(P<0.05);单细胞凝胶电泳显示实验组细胞DNA受损,浓度为25μmol/L、50μmol/L、100μmol/LPDTC处理后,实验组细胞DNA总损伤细胞百分率分别为43.50%、84.00%、95.63%,明显高于对照组(9.75%),P<0.05,且存在明显的剂量依赖关系;Western blotting结果显示经PDTC处理后的K562细胞胞质中可检测到pro-caspase-3和活化型caspase-3蛋白的表达。结论:NF-κB参与白血病细胞的增殖与凋亡调控,PDTC抗肿瘤机制可能与抑制NF-κB活性,上调caspase-3表达,从而诱导细胞凋亡有关。AIM： To investigate the effect of pyrrolidine dithiocarbamate （PDTC）, a specific inhibitor of NF - kB on the proliferation and apoptosis of K562 cells and to explore the anti - tumor mechanism of PDTC. METHODS： Trans AMTM NF - KB p65 kit was used to detect the activity of p65 in K562 cells treated by PDTC. The effect of PDTC on the proliferation of K562 cells was measured by WST - 1 method. I）NA damage was detected by single cell gel electrophoresis （ comet assay）. The procaspase - 3 and activated protein level of caspase - 3 were detected by Western blotting. RESULTS ： The activity of p65 in K.562 cells was inhibited after treated by PDTC （ P 〈 0.01 ）. Simultaneously the cell proliferation was significantly inhibited in a dose - and time - dependent manner （ P 〈 0.01 ）. The degree of DNA damage in K562 cells treated with PDTC at concentrations of 25 μmol/L, 50μmol/L or 100 μmol/L was more severe than that in control. The rates of comet cells in the PDTC - treated groups （43.50% , 84.00% , 95.63% ） were significantly higher than those in control （9.75%, P 〈0. 01 ）, and it was also dose - dependent. The expression of procaspase -3 and activated caspase -3 protein were detected in the cytoplasm of the K562 cells treated by PDTC by Western blotting. CONCLUSION ： NF -kB plays an important role in regulating cell proliferation and apoptosis in K562 cells. PDTC inhibits NF -kB activity and elevates the expression of caspase - 3, which is related to increase in cell apoptosis.

关 键 词：NF-ΚB抑制剂 细胞增殖 细胞凋亡 K562细胞 吡咯烷二硫代氨基甲酸盐 

分 类 号：R733.7[医药卫生—肿瘤]