论文部分内容阅读
以小鼠白血病细胞系L12 10为对象 ,探讨了抗白血病药物三尖杉酯碱 (Harringtonine,HT或Har)对细胞内着丝粒蛋白含量及着丝粒蛋白CenpB基因表达的影响。间接免疫荧光 (IIF)检测结果显示 ,随HT作用时间延长 ,L12 10细胞着丝粒荧光斑点减弱 ;免疫印迹 (Westernblot)检测结果显示 ,所用的抗着丝粒抗血清 (ACA血清 )能够识别 8种不同分子量的着丝粒蛋白 :14 0、80、70、5 6、37、34、32和 17kD。受HT作用 ,细胞中这些着丝粒蛋白的含量不同程度地降低。在L12 10细胞中识别 17、80and 14 0kD蛋白质的ACA抗体也分别与分子量相当的已知为CenpA、CenpB和CenpC的 3种蛋白发生交叉反应。Northern和Dotblot显示 ,HT的抑制作用使细胞中CenpBmRNA表达水平较之对照细胞下降。结果表明 ,HT可 (通过抑制基因mRNA的表达 )降低细胞中某些着丝粒蛋白的含量 ;HT对细胞的杀伤及诱导凋亡作用可能与CenpB等着丝粒蛋白基因的表达抑制有关
To investigate the effect of anti-leukemia drug harringtonine (HT or Har) on the expression of centromere protein and CenpB gene in murine leukemia cell line L1210. Indirect immunofluorescence (IIF) test results showed that with the prolongation of HT time, the centromere fluorescence spots of L12 10 cells were weakened. Western blot showed that the anti-centromeric antiserum (ACA serum) used could recognize 8 Centromere proteins of different molecular weights: 140, 80, 70, 56, 37, 34, 32 and 17 kD. Affected by HT, the levels of these centromere proteins in the cells are reduced to varying degrees. ACA antibodies that recognize 17,80 and 140 kD proteins in L12 10 cells also cross-react with three proteins of comparable molecular weight, known as CenpA, CenpB and CenpC, respectively. Northern and Dotblot showed that inhibition of HT decreased the level of CenpB mRNA expression in cells compared to control cells. The results showed that HT could decrease some centromere protein contents (by inhibiting gene mRNA expression). The cytotoxicity and apoptosis induction by HT could be related to the inhibition of CenpB isoform expression