塞来昔布联合三氧化二砷对慢性粒细胞白血病原代细胞bcr—abl蛋白及信号转导的影响  

Effects of celecoxib combined with arsenic trioxide on bcr-abl protein and signal transduction in CML primary cells

作　　者：刘定胜[1] 李玉峰[1] 李敏[2] 曹维克[3] 朱家斌[1] 何正梅[1] 

机构地区：[1]南京医科大学附属淮安第一医院血液科,223300 [2]南京医科大学附属淮安第一医院感染科,223300 [3]南京医科大学附属淮安第一医院中心实验室,223300

出　　处：《白血病．淋巴瘤》2011年第12期738-741,共4页Journal of Leukemia & Lymphoma

基　　金：南京医科大学科技发展基金（07NMUZ039）;淮安市科技发展计划（HAS07034）

摘　　要：目的探讨塞来昔布联合三氧化二砷（As2O3）对慢性粒细胞白血病（CML）原代细胞bcr-abl融合基因mRNA表达、bcr-abl蛋白即p210蛋白表达、蛋白质酪氨激酶（PTK）活性以及对下游增殖信号转导途径的影响。方法用塞来昔布（40μmol／L）、As2O3（2μmol／L）单独以及二者联合干预CML患者骨髓单个核细胞36h，进行实时荧光定量反转录聚合酶链反应（RQ-RT-PCR）、Western印迹和PTK活性检测，分析bcr—abl融合基因mRNA的表达、p210蛋白表达以及PTK活性的变化，以Western印迹检测STATl、STAT5、磷酸化STAT1（P-STAT1）、磷酸化STAT5（p-STAT5）以及IDl的表达变化。结果RQ-RT-PCR结果显示，对照组、塞来昔布组、As2O3组和塞来昔布联合As2O3组的mRNA水平分别为（5．97±0．53）％、（5．74±0．46）％、（5．94±O．57）％和（3．06±0．41）％，仅塞来昔布联合As203组与对照组的比较差异有统计学意义（t=28．35，P=0．00）。p210蛋白表达量化分析显示，塞来昔布与Asn均可下调p210蛋白，而塞来昔布联合As20。则显著下调p210蛋白。PTK活性检测显示，对照组、塞来昔布组、As203组和塞来昔布联合As203组吸光度（A450）值分别为1｜17±0．11、1．14±0．19、1．16±0．21和0．83±0．15，塞来昔布联合As2O3组与对照组比较差异有统计学意义（t=4．38，P=0．04）。Western印迹检测显示，塞来昔布联合As2O3可更显著下调STATl、STAT5、p-STATl、P—STATS和IDI蛋白表达。结论塞来昔布联合As，0，对下调CML原代细胞bcr-ablmRNA和蛋白表达、抑制PTK活性、抑制STAT及IDl信号转导通路具有协同作用。Objective To explore the effects of celecoxib combined with arsenic trioxide （As203） on the mRNA expression, protein expression and protein tyrosine kinase （PTK） activity of bcr-abl fusion gene and its downstream signal transduction in chronic myeloid leukemia （CML） primary cells. Methods The ceils were incubated with celecoxib （40 μmol/L） or As203 （2 μmol/L） alone and celecoxib （40 μmol/L） combined with As203 （2 μmol/L） for 36 hours. The changes of mRNA expression, p210 expression and PTK activity of bcr-abl fusion gene in each group were examined respectively by real time quantitative reverse transcriptase polymerase chain reaction （RQ-RT-PCR）, Western blot and PTK activity analysis. The important proteins STAT1, STAT5 and their phosphorylatic proteins p-STAT1 and p-STAT5 and inhibitor of DNA binding 1 （ID1） a common downstream target of oncogenic tyrosine kinase were also analyzed by Western blot. Results The mRNA expressions in control group, celecoxib group, As203 group and celecoxib combined with As203 group were （5.97±0.53） %, （5.74±0.46） %, （5.94±0.57） % and （3.06±0.41） % respectively, and the statistical difference was found only between celeeoxib combined with As203 group and control group （t = 28.35, P =0.00）. The similar statistic difference was only observed between the two groups from PTK activity tests （t = 4.38, P =0.04）. Western blot also showed that p210, STAT1, STAT5, p-STAT1, p-STAT5 and ID1 were more extraordinaryly downregulated by celecoxib combined with AszO3 than others treatments. Conclusion Celecoxib combined with As203 can downregulate mRNA, p210 expression, PTK activity of ber-abl fusion gene and inhibit STAT and ID1 signal transduction pathways in a synergistic way.

关 键 词：白血病 髓样 融合蛋白类 bcr—abl 塞来昔布 三氧化二砷 

分 类 号：R733.72[医药卫生—肿瘤]