高三尖杉酯碱联合AG490对HEL细胞JAK2-STAT5相关信号通路的影响  

Effect of Homoharringtonine Combined with AG490 on JAK2-STAT5 Associated Signal Pathway in HEL Cells

作　　者：任艳玲[1] 佟红艳[2] 

机构地区：[1]浙江省立同德医院血液科,浙江杭州310012 [2]浙江大学附属第一医院血液病研究所,浙江杭州310007

出　　处：《中国实验血液学杂志》2011年第5期1117-1120,共4页Journal of Experimental Hematology

摘　　要：本研究旨在探讨高三尖杉酯碱(homoharringtonine,HHT)联合AG490对HEL细胞JAK2-STAT5信号通路的影响及其机制,为临床应用新方案治疗慢性骨髓增殖性肿瘤(MPN)提供理论依据。以20 ng/ml的HHT,100μmol/L AG490,20 ng/ml HHT+100μmol/L AG490处理HEL细胞24、48、72小时以后,用MTT法和流式细胞术检测细胞生长抑制率和凋亡率,药物处理细胞24小时后用WB法检测JAK2突变激活的信号蛋白P-JAK2,P-STAT5以及BCL-xL表达的变化。结果表明,HHT以及AG490作用HEL 24小时均能抑制其增长,Annexin V-PI双染流式细胞图显示均能明显诱导HEL早期凋亡,HHT更为明显;免疫电泳分析显示,P-JAK2和P-STAT5表达下调,而JAK2和STAT5总蛋白水平稳定。结论:HHT联合AG490能明显抑制HEL细胞增殖并诱导凋亡,两者具有协同作用,其作用机制是HHT作为一种广谱的蛋白酪氨酸激酶抑制剂,协同AG490抑制JAK2突变引起的信号蛋白的酪氨酸位点的磷酸化,从而下调STAT5反应性基因的转录。This study was aimed to explore the effect of homoharringtonine in combination with AG490 on JAK2-STAT5 associated signal pathway in HEL cells,and analyze its mechanism so as to provide theoretical basis for therapy of chronic myeloproliferative neoplasma by new program.The cell survival rates were tested by MTT,apoptosis was tested by flow cytometry after HEL cells were treated by 20 ng/ml HHT,100 μmol/L AG490 and 20 ng/ml HHT in combination with 100 μmol/L AG490,while the signal proteins such as P-JAK2,P-STAT5 and BCL-xL activated by abnormal activated JAK2 were tested by Western blot.The results showed that both HHT and AG490 could inhabit the HEL cell proliferation after being treated for 24 hours,and Annexin V-PI double staining confirmed early apoptosis while HHT effect was more obvious,Western blot showed that the expressions of P-JAK2 and P-STAT5 were down-regulated,while the total protein levels of JAK2 and STAT5 were stable.It is concluded that HHT combined with AG490 can obviously inhibit the proliferation and induce early apoptosis of HEL cells,and there is synergistic effect between the two drugs.HHT possibly acts as a broad-spectrum PTK inhibitor and synergistically with AG490 inhibits the phosphorylation of signal proteins caused by JAK2V617F,thus down-regulating the transcription of STAT5.

关 键 词：高三尖杉酯碱 AG490 HEL细胞 JAK2-STAT5信号通路 慢性骨髓增殖性肿瘤 

分 类 号：R551.3[医药卫生—血液循环系统疾病]