Autophagy governs erythroid differentiation both in vitro and in vivo
Autophagy governs erythroid differentiation both in vitro and in vivo
复制标题
自噬在体外和体内控制红细胞分化
DOI：
10.1179/1607845415y.0000000027
复制
发表时间：
2016-01-01
期刊：
影响因子：
1.9
通讯作者：
Zhang, Suping
中科院分区：
文献类型：
--
作者：
Cao, Yan;Cai, Jinyang;Zhang, Suping
目的：尽管近年来自噬在决定细胞命运中的重要性已被广泛探讨，但其在调节红细胞分化中的确切作用仍不清楚。方法：以人红细胞白血病细胞系K562为细胞模型，在体外研究红细胞分化。用饥饿和雷帕霉素诱导自噬，用Baf-A1和CRISPR/Cas9/Atg7分别抑制晚期和早期自噬。建立小鼠自噬激活和自噬缺失模型，利用红细胞计数和流式细胞术分析红细胞在体内的分化情况。结果：经自噬抑制剂、Baf-A1或Atg7基因敲除后，经饥饿和雷帕霉素处理的K562细胞α -珠蛋白和γ -珠蛋白转录水平及血红蛋白阳性细胞比例均显著升高。在自噬增强小鼠模型中，成熟红细胞数量明显增加，而在atg7缺失小鼠模型中，红细胞分化严重受阻。结论：无论在体内还是体外，红细胞的发育、成熟和稳态都离不开自噬。我们的研究结果支持了通过调节自噬来治疗红细胞白血病和在体外产生红细胞的潜在策略。
Objectives: Although the importance of autophagy in determination of cell fate has been much explored in recent years, its definite role in regulating erythroid differentiation remains unclear.Methods: In this study, human erythroleukemic cell line K562 was employed as a cell model for studying erythroid differentiation in vitro. Starvation and rapamycin were used to induce autophagy, whereas Baf-A1 and CRISPR/Cas9/Atg7 were used to inhibit late and early phase of autophagy, respectively. The mice model of autophagy activation and autophagy deletion were established, and red blood cell counts and flow cytometry were used to analyze erythroid differentiation in vivo.Results: The results showed that the transcriptional levels of alpha-globin and gamma-globin, and the ratio of hemoglobin-positive cells all significantly increased in K562 cells with starvation and rapamycin treatment, which were blocked by autophagy inhibitor, Baf-A1 or Atg7 gene knockout. In the autophagy-enhanced mouse model, the number of mature erythrocytes significantly increased, while in Atg7-deleted mouse model, erythroid differentiation was severely blocked.Conclusion: It is concluded that autophagy is indispensible in the development, maturation, and homeostasis of erythroid cells both in vitro and in vivo. Our findings support the potential strategy for erythroleukemia treatment and production of erythroblasts in vitro by modulating autophagy.