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目的探讨当归多糖(APS)诱导K562细胞向红系样细胞分化相关的信号转导通路。方法实验分组:对照组采用常规方法培养;APS组在常规培养基础上加入APS(终浓度100～500mg/L),分别培养12h、1d、2d、3d。联苯胺染色与分光光度法检测经APS诱导后K562细胞向红系样细胞分化的血红蛋白表达;激光扫描共焦显微镜观察JAK2、信号转导和转录激活因子5(STAT5)在各组细胞内的分布;Westernblotting检测细胞核、质蛋白中JAK2、STAT5的表达变化;免疫沉淀法检测质蛋白中JAK2的磷酸化改变。结果经APS诱导后K562细胞的联苯胺染色阳性率增高,随着APS诱导浓度增加K562细胞合成血红蛋白也增加;APS诱导K562细胞12h、24h和48h,核蛋白中STAT5表达较对照组增加,质蛋白中STAT5表达减少;APS组与对照组K562细胞的JAK2表达未见显著差异,但APS组的JAK2磷酸化强于对照组。结论APS可诱导K562细胞向红系样细胞分化,其机制可能与APS启动JAK2的酪氨酸磷酸化,继而引起STAT5核转位,通过信号转导通路的调控影响细胞的增殖分化。
Objective To investigate the signal transduction pathways involved in the differentiation of K562 cells into erythroid-like cells induced by Angelica sinensis polysaccharide (APS). Methods Experimental group: control group were cultured by conventional methods; APS group were added APS (final concentration 100 ~ 500mg / L) on the basis of routine culture and cultured for 12h, 1d, 2d and 3d respectively. Benzidine staining and spectrophotometry were used to detect the hemoglobin expression of K562 cells differentiated to erythroid-like cells induced by APS. The distribution of JAK2, signal transducers and activators of transcription 5 (STAT5) in each group was observed by laser scanning confocal microscope Western blotting was used to detect the expression of JAK2 and STAT5 in the nuclei and proteome. The phosphorylation of JAK2 was detected by immunoprecipitation. Results The positive rate of benzidine staining of K562 cells induced by APS was increased. The hemoglobin synthesis of K562 cells increased with the increase of APS concentration. The expression of STAT5 in nucleoside of K562 cells induced by APS was increased at 12h, 24h and 48h But there was no significant difference in JAK2 expression in K562 cells between APS group and control group. However, JAK2 phosphorylation in APS group was stronger than that in control group. Conclusion APS can induce the differentiation of K562 cells into erythroid lineage-like cells. The mechanism may be related to the tyrosine phosphorylation of JAK2 initiated by APS and the nuclear translocation of STAT5. APS can influence the proliferation and differentiation of cells through the regulation of signal transduction pathway.