黄色黄素抑制人白血病HL-60细胞内蛋白激酶CK2的实验研究  

The Study of Digitoflavone for The Inhibition of Protein Kinase CK2 in HL-60 Cells

作　　者：林小聪[1] 刘新光[1] 阮杰[1] 陈小文[1] 梁念慈[1] 

机构地区：[1]广东医学院生物化学与分子生物学研究所,湛江524023

出　　处：《生物化学与生物物理进展》2007年第2期187-195,共9页Progress In Biochemistry and Biophysics

基　　金：教育部科学技术研究重点项目(03096)~~

摘　　要：蛋白激酶CK2在寻找抗肿瘤及抗病毒药物方面是一个具有吸引力的分子靶点,其特异性抑制剂具有潜在的临床应用价值.通过测定药物作用后转移到CK2底物上[γ-32P]ATP的放射性活度,探讨黄色黄素对重组人CK2全酶以及细胞内CK2活性的影响;采用多重RT-PCR检测CK2α、α'和β亚基的mRNA表达水平;Lineweaver-Burk作图法分析CK2的酶动力学机制.发现黄色黄素能显著抑制重组人CK2全酶(IC50=0.86μmol/L)以及HL-60细胞内的CK2活性,其作用效果均强于阳性对照TBB.另外,黄色黄素作用2h可使CK2α和α'亚基的mRNA表达下降,对β亚基则无明显的影响.酶动力学分析表明,黄色黄素与ATP呈竞争性抑制CK2的活性,与酪蛋白则呈混合性抑制CK2的活性.研究说明,黄色黄素是一种有效的细胞内蛋白激酶CK2抑制剂.Protein kinase CK2 is an attractive target for anti-neoplastic and antiviral drugs and the inhibitors of CK2 have clinical therapeutic potential. The effect of recombinant human CK2 holoenzyme activity and cellular CK2 activity by digitoflavone was assayed by detecting incorporation of 32p of [γ-^32p]ATP into the substrate. The mRNA expression of CK2α,α＇ and β subunits were detected by multiplex RT-PCR. CK2 kinetic analysis was carried out by using the Lineweaver-Burk plot. Digitoflavone was shown to inhibit strongly the activity of recombinant human CK2 holoenzyme in vitro （IC50=0.86μmol/L） and endogenous CK2 in HL-60 cells, which was more effective than the positive control 4Br-2-azabenzimidazole （TBB）. After being treated for 2 h with digitoflavone, the mRNA expression of CK2α and α＇ subunit decreased, while β subunit was not changed fundamentally. Kinetic studies of digitoflavone on recombinant human CK2 showed that digitoflavone acted as an inhibitor of competitive with ATP and mixed types with casein. These results indicating that digitoflavone is a potent endogenous inhibitor of protein kinase CK2.

关 键 词：蛋白激酶CK2 黄色黄素 HL-60细胞 

分 类 号：Q555.7[生物学—生物化学] Q814.9