伴CSF1R-Y571D突变的急性髓系白血病细胞系GDM-1的增殖机制  

Proliferation mechanism of acute myeloid leukemia cell line GDM-1 with CSF1R-Y571D mutation

作　　者：曹琼 徐凝馨 李小文 吴昊 郝建卿 覃艳红 李莉[1] CAO Qiong;XU Ning-xin;LI Xiao-wen;WU Hao;HAO Jian-qing;QIN Yan-hong;LI Li(School of Basic Medicine, Shanxi Medical University, Taiyuan 030001;Department of Hematology, the Second Hospital of Shanxi Medical University, Taiyuan 030001, China)

机构地区：[1]山西医科大学基础医学院,山西太原030001 [2]山西医科大学第二临床医学院血液科,山西太原030001

出　　处：《基础医学与临床》2021年第1期44-49,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金青年科学基金(81400139);山西省自然科学基金(2014011039-2)。

摘　　要：目的通过蛋白激酶抑制方法研究伴集落刺激因子-1受体(CSF1R)突变的急性髓系白血病细胞系GDM-1的增殖机制。方法按照处理方式不同将GDM-1细胞分为实验组和对照组,将10种不同的蛋白激酶抑制剂作用于实验组,对照组仅做DMSO处理;CCK8法检测细胞增殖抑制情况;Western blot检测蛋白激酶抑制剂作用于GDM-1后,关键激酶的磷酸化水平的改变;细胞因子M-CSF单独或与蛋白激酶抑制剂共同作用于GDM-1细胞,采用CCK8法进行检测细胞增殖水平。结果蛋白激酶抑制分析显示明显抑制GDM-1细胞增殖的药物有dasatinib(IC50=0.01μmol/L)、bosutinib(IC50=0.07μmol/L)、linifanib(IC50=0.13μmol/L)及sorafenib(IC50=0.12μmol/L)。Western blot显示,与对照组相比,linifanib作用于GDM-1后,SRC、ERK磷酸化水平明显降低(P<0.01)。在低浓度FBS培养时,细胞因子M-CSF能够刺激伴CSF1R突变型细胞系GDM-1的增殖,其增殖水平为对照组的11.3倍;M-CSF联合bosutinib作用于GDM-1,细胞增殖明显被抑制(P<0.001),而M-CSF联合HG6-64-1作用于GDM-1,与仅用M-CSF组相比细胞增殖不受影响。结论GDM-1细胞中CSF1R-Y571D突变使CSF1R活性增高,并主要通过激活Src及MAPK信号通路来促进细胞增殖。Objective To explore the proliferation mechanism of acute myeloid leukemia cell line GDM-1 by the method of protein kinase inhibition.Methods GDM-1 cells were divided into experimental group and control group according to different treatment.Experimental group was treated with 10 different protein kinase inhibitors,the control group was treated with DMSO,and the inhibition of cell proliferation was detected by CCK8 method.The phosphorylation level of key protein kinases in GDM-1 after treatment with linifanib,bosutinib,sorafenib were detected by Western blot.GDM-1 cells were treated with cytokine M-CSF alone or combined with protein kinase inhibitors,cell proliferation was detected by CCK8 method.Results Protein kinase inhibition analysis showed that dasatinib(IC50=0.01μmol/L),bosutinib(IC50=0.07μmol/L),linifanib(IC50=0.13μmol/L)and sorafenib(IC50=0.12μmol/L)significantly inhibited the proliferation of GDM-1 cells.Western blot analysis showed that the phosphorylation level of Src and ERK in GDM-1 was decreased(P<0.01)when treated with linifaninb.The proliferation of GDM-1 was 11.3 times as much as that of control group when stimulated with M-CSF.The proliferation of GDM-1 was dramatically inhibited when incubated with the combination of M-CSF and bosutinib(P<0.001),but it was not significantly inhibited when incubated with M-CSF plus HG6-64-1.Conclusions CSF1R-Y571D mutation of GDM-1 cells increases the activity of CSF1R,and promotes cell proliferation by activating Src and MAPK signaling pathways.

关 键 词：蛋白激酶抑制剂 GDM-1 集落刺激因子1受体 linifanib BOSUTINIB 

分 类 号：R733.71[医药卫生—肿瘤]