Wnt-10a在氯高铁血红素诱导K562细胞分化过程中的作用

Functions of Wnt-10a during K562 cell differentiation induced by hemin

目的探讨Wnt-10a在氯高铁血红素(hemin)诱导K562细胞分化过程中的变化及功能。方法通过联苯胺染色检测K562细胞被hemin诱导的阳性率,利用Western blotting和免疫细胞化学分析K562细胞诱导分化过程中Wnt-10a表达水平和细胞定位的变化情况,通过半定量PCR与Real-time PCR检测K562细胞诱导分化过程中Wnt信号途径关键因子的表达水平及其变化,通过Wnt信号途径激活剂与构建Wnt-10a高表达K562细胞株的方法探讨Wnt途径改变后对K562细胞诱导分化的影响。结果在hemin诱导K562细胞分化过程中,细胞中的Wnt-10a蛋白表达量和mRNA量都出现了短暂下降后回复的趋势,同时Wnt-10a向胞膜迁移,Wnt蛋白所参与的3条信号途径中关键因子的表达水平均有着不同程度的变化。使用Wnt信号通路抑制剂后,诱导分化过程中的细胞增殖活力加强,Wnt-10a高表达的K562细胞被hemin诱导分化的能力开始提高,并且Wnt经典信号途径的关键因子表达水平也有上升。结论 Wnt-10a与hemin诱导K562细胞红系分化的过程密切相关。

Objective To study expression changes and functions of Wnt-10a protein in K562 differentiation indueed by heroin. Methods Benzidine staining was used to test hemin-induced erythroid differentiation of K562 cells. Western blotting and immunocytoehemistry were used to detect protein expression ehanges of Wnt-10a and protein localization in K562 cells. Real-time PCR and Real-time PCR were applied to investigate mRNA expression of key proteins in Wnt signalling pathway during K562 defferentiation process. Wnt signaling pathway activator and Wnt-10a high expression K562 cell line were used to explore the influence of altered Wnt pathway. Results Wnt-10a protein and mRNA expressions were upregulated after a brief down in the erythroid differentiation proeess of K562 cells induced by hemin when, Wnt-10a protein migrated to the cell membrane. The key factors of the main Wnt signaling pathways changed during K562 differentiation. By using Wnt signaling pathway inhihitor, cell proliferation capability was enhanced during K562 differentiation by heroin. When Wnt-10a was highly expressed, the ability of induced differentiation of K562 cells was improved. Conclusion These results show that Wnt-10a protein is closely related with K562 cell differentiation induced by heroin, which may offer clues for further study of the mechanism of leukemia and cell differentiation.