Fan H, Kanji S, Zhai M
Department of Hematology, The First Clinical College, China Medical University, Shenyang 110001, China.
Zhonghua Xue Ye Xue Za Zhi. 2000 Jun;21(6):294-7.
To investigate the expression of erythropoietin receptor (EpoR) in leukemic cell lines and to clarify the mechanism of proliferative signal transmission in the leukemic cell line KOCL-33 mediated by EpoR.
Biotinylated Epo and flow cytometry were used for EpoR expression, (3)H-TdR incorporation for cell proliferation, immunoprecipitation and Western blot analysis for the tyrosine phosphorylation of signal transmission proteins.
(1) All 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. (2) The 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. (3) The 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.
(1) There was EpoR expression in leukemic cell lines. (2) The 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. (3) Dimerization and tyrosine phosphorylation of EpoR occurred, and JAK2, STAT5 involved in the proliferative signal transmission in KOCL-33 cells mediated by EpoR.
研究促红细胞生成素受体(EpoR)在白血病细胞系中的表达情况,并阐明EpoR介导的白血病细胞系KOCL-33增殖信号转导机制。
采用生物素化促红细胞生成素和流式细胞术检测EpoR表达,用³H-TdR掺入法检测细胞增殖,用免疫沉淀和蛋白质印迹分析检测信号转导蛋白的酪氨酸磷酸化。
(1)除T淋巴细胞系外,所有检测的细胞系均表达EpoR。阳性率为18%~99%,平均为52%,T、B和非淋巴细胞之间无差异。(2)在9个检测的细胞系中,有7个细胞系对促红细胞生成素的反应是细胞增殖显著增强。促红细胞生成素的刺激作用与细胞上表达的促红细胞生成素受体密度无关。(3)促红细胞生成素刺激后,KOCL-33细胞的增殖显著增强。促红细胞生成素刺激后1分钟,EpoR发生二聚化和酪氨酸磷酸化,JAK2发生酪氨酸磷酸化;5分钟后,STAT5发生酪氨酸磷酸化。
(1)白血病细胞系中有EpoR表达。(2)部分白血病细胞系中促红细胞生成素可刺激细胞增殖,促红细胞生成素的刺激作用与细胞上表达的促红细胞生成素受体密度无关。(3)EpoR发生二聚化和酪氨酸磷酸化,JAK2、STAT5参与EpoR介导的KOCL-33细胞增殖信号转导。