摘要
目的 检测白血病细胞系红细胞生成素受体 (EpoR)的表达并阐明EpoR介导的白血病细胞系KOCL 33增殖信号传导途径。方法 用生物素酰基化Epo及流式细胞仪检测白血病细胞系EpoR的表达 ;3H TdR掺入法检测各细胞系对Epo刺激的增殖反应 ;免疫沉淀法和Western印迹法检测KOCL 33细胞中几种信号传导蛋白的酪氨酸磷酸化。结果 ①除T淋巴细胞系外 ,其余细胞系EpoR表达均为阳性 ,阳性率为 18%～ 99% ,均值 5 2 %。不同类型的细胞系EpoR阳性率的差异没有统计学意义。② 9株细胞中 7株细胞因受刺激而显著地增殖 ,P值 <0 .0 5。EpoR阳性率与增殖反应不相一致。③Epo可引起KOCL 33细胞的增殖 ,刺激后 1min ,EpoR形成同源二聚体、出现酪氨酸磷酸化 ,JAK2酪氨酸磷酸化 ;5min时STAT5被酪氨酸磷酸化。结论 ①白血病细胞系可有EpoR的表达 ;②Epo可引起部分白血病细胞系的增殖 ;③在EpoR介导的KOCL 33细胞增殖信号的传导中 ,EpoR形成同源二聚体、出现酪氨酸磷酸化 ,JAK2、STAT5参与了增殖信号的传导。
Objective To investigate the expression of erythropoietin receptor (EpoR) in leukemic cell lines and to clearify the mechanism of proliferative signal transmission in the leukemic cell line KOCL 33 mediated by EpoR. Methods Biotinylated Epo and flow cytometry were used for EpoR expression, 3H TdR incorporation for cell proliferation, immunoprecipitation and Western blot analysis for the tyrosine phosphorylation of signal transmission proteins. Results ①All the cell lines tested expressed EpoR except for T lymphoid cell lines. The positivity rates ranged from 18% to 99% with an average of 52%, and was no difference among T, B and non lymphoid cells. ②The cell proliferation was significantly enhanced in response to Epo in 7 of 9 cell lines tested. The stimulating effects of Epo did not correlate with the densities of EpoR expressed on the cells. ③The proliferation of KOCL 33 cells was significantly enhanced in response to Epo. Dimerization and tyrosine phosphorylation of EpoR and tyrosine phosphorylation of JAK2 occurred in 1?min and tyrosine phosphorylation of STAT5 in 5 minutes after Epo stimulation. Conclusion ①There was EpoR expression in leukemic cell lines. ②The cell proliferation could be stimulated by Epo in some leukemic cell lines. The stimulating effect of Epo did not correlate with the densities of EpoR expressed on the cells.③Dimerization and tyrosine phosphorylation of EpoR occurred,and JAK2, STAT5 involved in the proliferative signal transmission in KOCL 33 cells mediated by EpoR.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2000年第6期294-297,共4页
Chinese Journal of Hematology
关键词
白血病
细胞系
增殖信号传导
红细胞生成素
受体
Cell line
Receptor, erythropoietin(EpoR)
Signal transduction
Tyrosine phosphorylation
