Jagged-1抑制小鼠胚胎干细胞分化为造血干/祖细胞

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (1): 55-61.

Jagged-1抑制小鼠胚胎干细胞分化为造血干/祖细胞

陈日玲¹, 郑伟荣², 谭霖³, 刘东强³, 史惠³, 陈启康^1*

1.广东医科大学附属顺德妇女儿童医院儿科, 广东顺德 528300;
2.泉州市第一医院儿科, 福建泉州 362000;
3.广东医科大学附属医院儿童医学中心, 广东湛江 524023

收稿日期:2019-05-16 修回日期:2020-03-30 出版日期:2021-01-05 发布日期:2020-12-30
通讯作者: ^*chen.qikang@163.com
基金资助:
湛江市科技竞争性项目(2014A01022)

Jagged-1 inhibits mouse embryonic stem cells differentiating into haematopoietic stem or progenitor cells

CHEN Ri-lin¹, ZHENG Wei-rong², TAN Lin³, LIU Dong-qiang³, SHI Hui³, CHEN Qi-kang^1*

1. Department of Pediatrics,the Affiliated Shunde Women and Children Hospital, Guangdong Medical University, Shunde 528300;
2. Department of Pediatrics, the First Hospital of Quanzhou,Quanzhou 362000;
3. Children's Medical Center, the Affiliated Hospital of Guangdong Medical University, Zhanjiang 524023, China

Received:2019-05-16 Revised:2020-03-30 Online:2021-01-05 Published:2020-12-30
Contact: ^*chen.qikang@163.com

摘要/Abstract

摘要： 目的探讨Notch信号通路在小鼠胚胎干细胞(ESC)分化为造血干/祖细胞(HSC/HPC)中的作用。方法 1)体外培养小鼠拟胚体细胞(EBs),使用Jagged-1蛋白活化Notch信号通路及DAPT(γ-分泌酶抑制剂)抑制Notch信号通路。实验分为拟胚体细胞复苏组(EB组)、对照组、Jagged-1组、DAPT组和Jagged-1-DAPT组。2)流式细胞计量术检测ESC特异性表型和HSC/HPC特异性表型的表达。3)实时定量PCR检测各组Notch信号通路基因、小鼠ESC表型基因和HSC/HPC表型基因的表达。结果 1) Jagged-1组细胞胚胎干细胞数较对照组和Jagged-1-DAPT组明显增多(P＜0.05), 2)Jagged-1-DAPT组分化的HSC/HPC数较control组及Jagged-1组明显增多(P＜0.05);3)Jagged-1组Notch1、Notch2、Notch4 mRNA表达量较对照组明显升高(P＜0.05);4)DAPT组和Jagged-1-DAPT组中Notch1、Notch4 mRNA表达量较对照组降低(P＜0.05)。结论 Jagged-1激活Notch信号通路可抑制小鼠ESC向HSC/HPC的分化。

关键词: 胚胎干细胞, Notch信号通路, 造血干细胞, Jagged-1

Abstract: Objective To investigate the effect of Notch signaling pathways on differentiation of mouse embryonic stem cells(ESC) into haematopoietic stem or progenitor cells(HSC/HPC). Methods 1)incubate mouse embryoid body cells and divide the embryonic stem cells into five groups as EB group,control group,Jagged-1 group, DAPT group and Jagged-1-DAPT group. 2)detect the phenotype of HSC/HPC by flow cytometry. 3)The expression of Notch signaling pathway, mouse embryonic stem cell phenotype and HSC/HPC phenotype were detected by RT-PCR. Results 1)the number of embryonic stem cells in Jagged-1 group was significantly more than that in control group and Jagged-DAPT group(P＜0.05);2)the number of HSC/HPC in Jagged-1-DAPT group was significantly more than that in control group and in Jagged-1 group(P＜0.05); 3)the expression of Notch1, Notch2 and Notch4 mRNA in Jagged-1 group was significantly higher than that in control group(P＜0.05), 4)the expression of Notch1 and Notch4 mRNA in DAPT group and Jagged-1-DAPT group was lower than that in control group(P＜0.05). Conclusions Jagged-1 activation of Notch signaling pathway potentially inhibits ESC differentiation into HSC/HPC in mice model.

Key words: embryonic stem cells, Notch signal pathway, hematopoietic stem cells, Jagged-1

中图分类号:

R329.28
Q253

陈日玲, 郑伟荣, 谭霖, 刘东强, 史惠, 陈启康. Jagged-1抑制小鼠胚胎干细胞分化为造血干/祖细胞[J]. 基础医学与临床, 2021, 41(1): 55-61.

CHEN Ri-lin, ZHENG Wei-rong, TAN Lin, LIU Dong-qiang, SHI Hui, CHEN Qi-kang. Jagged-1 inhibits mouse embryonic stem cells differentiating into haematopoietic stem or progenitor cells[J]. Basic & Clinical Medicine, 2021, 41(1): 55-61.

参考文献

[1] Dzierzak E, Bigas A. Blood development: hematopoietic stem cell dependence and independence[J]. Cell Stem Cell, 2018, 22: 639-651.
[2] Leung A, Zulick E, Skvir N, et al. Notch and aryl hydrocarbon receptor signaling impact definitive hematopoiesis from human pluripotent stem cells[J]. Stem Cells, 2018, 36:1004-1019.
[3] 宋波,陈日玲,廖凤玲,等. Notch 信号通路对VEGF促小鼠胚胎干细胞分化成为造血干/祖细胞的作用[J]. 中国实验血液学杂志, 2018,26:1-5
[4] Yamaguchi E, Chiba S, Kumano K, et al. Expression of Notch ligands, Jagged1, 2 and Delta1 in antigen presenting cells in mice.[J]. Immunol Lett, 2002, 81:59-64.
[5] Hao B, Chen X, Cao Y. Yes associated protein 1 pro-motes the metastasis of U251 glioma cells by upregulating Jagged-1 expression and activating the Notch signal pathway[J]. Exp Ther Med, 2018, 16: 1411-1416.
[6] 黄清昕,涂三芳,黄睿,等. 异基因造血干细胞移植治疗100例白血病的临床总结[J]. 中国实验血液学杂志,2016,24:556-561.
[7] Nair V, Apte S. Stem cell transplantation in aplastic anaemia[J]. Assoc Physi India, 2015, 63: 21-25.
[8] 刘恒铫, 赵国丽, 王福文,等. Notch信号通路在肺癌和肺癌干细胞中的作用及其抑制剂[J]. 国际肿瘤学杂志, 2018:250-252
[9] Suresh S, Irvine AE. The notch signaling pathway in normal and malignant blood cell production[J]. J Cell Commun Signal, 2015, 9:5-13.
[10] Bigas A, Altri TD,Espinosa L. The Notch pathway in hematopoietic stem cells[J]. Curr Top Microbiol Immunol, 2012,360: 1-18.
[11] Moriyama M, Osawa M, Mak SS,et al. Notch signaling via Hes1, transcription factor maintains survival of melanoblasts and melanocyte stem cells[J]. Cell Biol, 2006, 173:333-339.
[12] Kobayashi T, Mizuno H, Imayoshi I, et al. The cyclic gene Hes1 contributes to diverse differentiation responses of embryonic stem cells[J]. Genes Dev, 2009, 23: 1870-1875
[13] Huang C, Jackson M, Samuel K, et al. Haematopoietic differentiation is inhibited when Notch activity is enhanced in FLK1+ mesoderm progenitors[J]. Stem Cell Res, 2013, 11: 1273-1287.
[14] Liao F, Tan L, Liu H, et al. Hematopoietic stem cell-derived exosomes promote hematopoietic differentiation of mouse embryonic stem cells in vitro via inhibiting the miR126/Notch1 pathway[J]. Acta Pharmacol Sinica, 2018, 39: 552-588.

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