[Effect of Rheb1 in the Development of Mouse Megakaryocyte-Erythroid Progenitor Cells]

Juan Gao; Shuang Yang; Yu-Xia Wang; Ya-Nan Gao; Ya-Jing Chu; Wei-Ping Yuan; Xiao-Min Wang

doi:10.19746/j.cnki.issn.1009-2137.2022.01.049

[Effect of Rheb1 in the Development of Mouse Megakaryocyte-Erythroid Progenitor Cells]

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2022 Feb;30(1):298-304. doi: 10.19746/j.cnki.issn.1009-2137.2022.01.049.

[Article in Chinese]

Authors

Juan Gao¹, Shuang Yang², Yu-Xia Wang², Ya-Nan Gao³, Ya-Jing Chu², Wei-Ping Yuan², Xiao-Min Wang²

Affiliations

¹ 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 Science, Tianjin Eye Institute, Clinical College of Ophthalmology, Tianjin Medical University, Nankai University Affiliated Eye Hospital, Tianjin 300020, China.
² 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.
³ Department of Pathology, Tianjin Medical University Cancer Institute and Hospital, Tianjin Medical University, Tianjin 300060, China.

PMID: 35123643
DOI: 10.19746/j.cnki.issn.1009-2137.2022.01.049

Abstract
in English, Chinese

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;Rheb1^fl/fl mice(Rheb1^Δ/Δ mice). Flow cytometry was used to detect the percentage of red blood cells in peripheral blood and erythroid cells in bone marrow in Vav1-Cre;Rheb1^fl/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. Real-time fluorescence quantification PCR was used to detect the relative expression of PU.1,GATA-1,GATA-2,CEBPα and CEBPβ of 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 wild-type 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.

题目: Rheb1在小鼠巨核-红系多能性干细胞发育中的作用.

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;Rheb1^fl/fl, Rheb1^Δ/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.

Keywords: Rheb1; Rheb1^fl/fl; Vav1-Cre; megakaryocyte-erythroid progenitor cells; mouse; rapamycin.

MeSH terms

Animals
Cell Differentiation
Erythrocytes
Flow Cytometry
Megakaryocyte-Erythroid Progenitor Cells*
Megakaryocytes
Mice
Signal Transduction*

Abstract in English, Chinese

MeSH terms

Abstract
in English, Chinese