| Rheb1在小鼠巨核-红系多能性干细胞发育中的作用 |
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| 引用本文： | 高娟,杨爽,王玉霞,高亚男,初雅婧,袁卫平,汪晓敏.Rheb1在小鼠巨核-红系多能性干细胞发育中的作用[J].中国实验血液学杂志,2022(1):298-304. |
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| 作者姓名： | 高娟 杨爽 王玉霞 高亚男 初雅婧 袁卫平 汪晓敏 |
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| 作者单位： | 实验血液学国家重点实验室;天津市眼科医院;天津医科大学肿瘤医院病理科 |
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| 基金项目： | 国家自然科学基金(项目编号:81870088,81700103,81470280,81300436,81770105)。 |
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| 摘 要： | 目的:研究Rheb1基因在小鼠巨核-红系多能性干细胞发育成熟中的作用及相关的机制。方法:利用Vav-Cre在造血系统中特异性敲除小鼠Rheb1(Vav1-Cre;Rheb1fl/fl,Rheb1Δ/Δ小鼠),采用流式细胞术检测Rheb1敲除组和对照组小鼠骨髓和外周血中红系细胞的比例;CFC克隆形成实验检测敲除组和对照组小鼠骨髓中巨核-红系多能性干细胞体外克隆形成能力;利用实时荧光定量PCR检测敲除组和对照组小鼠巨核-红系多能性干细胞中PU.1、GATA-1、GATA-2、CEBPα和CEBPβ的相对表达量;在培养基中加入雷帕霉素,检测野生型小鼠巨核-红系多能性干细胞体外克隆形成能力的变化。结果:Rheb1敲除后,小鼠骨髓中红系细胞发育受抑且应激能力减弱,小鼠骨髓中巨核-红系多能性干细胞体外克隆形成能力减弱,GATA-1的表达水平降低;雷帕霉素可以抑制野生型小鼠骨髓中巨核-红系多能性干细胞体外克隆的形成。结论:Rheb1基因在小鼠巨核-红系多能性干细胞发育中具有重要的调控作用,Rheb1可能通过mTOR信号通路调控小鼠巨核-红系多能性干细胞发育...
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| 关 键 词： | Rheb1 巨核-红系多能性干细胞 雷帕霉素 Vav1-Cre Rheb1fl/fl小鼠 |
Effect of Rheb1 in the Development of Mouse Megakaryocyte-Erythroid Progenitor Cells |
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| GAO Juan,YANG Shuang,WANG Yu-Xia,GAO Ya-Nan,CHU Ya-Jing,YUAN Wei-Ping,WANG Xiao-Min.Effect of Rheb1 in the Development of Mouse Megakaryocyte-Erythroid Progenitor Cells[J].Journal of Experimental Hematology,2022(1):298-304. |
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| Authors: | GAO Juan YANG Shuang WANG Yu-Xia GAO Ya-Nan CHU Ya-Jing YUAN Wei-Ping WANG Xiao-Min |
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| Institution: | (State Key Laboratory of Experimental Hematology,National Clinical Research Center for Hematological Disorders,Institute of Hematology and Blood Diseases Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Tianjin 300020,China;Tianjin Eye Hospital,Tianjin Key Laboratory of Ophthalmology and Visual Sciencey Tianjin Eye Institute,Clinical College of Ophthalmology,Tianjin Medical Universityy Nankai University Ajfiliated Eye Hospital,Tianjin 300020,China;Department of Pathology,Tianjin Medical University Cancer Institute and Hospital,Tianjin Medical University,Tianjin 300060,China) |
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| Abstract: | Objective:To investigate the effect of Rheb1 in the development of mouse megakaryocyte-erythroid progenitor cells and its related mechanism.Methods:Rheb1 was specifically knocked-out in the hematopoietic system of Vav1-Cre;Rheb1fl/fl mice(Rheb1Δ/Δ mice).Flow cytometry was used to detect the percentage of red blood cells in peripheral blood and erythroid cells in bone marrow in Vavl-Cre;Rheb1fl/fl mice and control mice.The CFC assay was used to detect the differentiation ability of Rheb1 KO megakaryocyte-erythroid progenitor cells and control cells.Realtime fluorescence quantification PCR was used to detect the relative expression of PU.1,GA TA-1,GATA-2,CEBPαand CEBPβof Rheb1 KO megakaryocyte-erythroid progenitor cells and control cells.Rapamycin was added to the culture medium,and it was used to detect the changes in cloning ability of megakaryocyte-erythroid progenitor cells from wildtype mice in vitro.Results:After Rheb1 was knocked out,the development and stress response ability of megakaryocyte-erythroid progenitor cells in mice were weaken and the differentiation ability of megakaryocyte-erythroid progenitor cells in vitro was weaken.Moreover,the expression of GATA-1 of megakaryocyte-erythroid progenitor cells was decreased.Further,rapamycin could inhibit the differentiative capacity of megakaryocyte-erythroid progenitor cells in vitro.Conclusion:Rheb1 can regulate the development of megakaryocyte-erythroid progenitor cells probably through the mTOR signaling pathway in mice. |
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| Keywords: | Rheb1 megakaryocyte-erythroid progenitor cells rapamycin Vav1-Cre Rheb1fl/fl mouse |
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