Uhrf1对肠上皮发育的影响

doi:10.16288/j.yczz.20-337

遗传 ›› 2021, Vol. 43 ›› Issue (1): 84-93.doi: 10.16288/j.yczz.20-337

Uhrf1对肠上皮发育的影响

王芯悦^1,³, 李亮^2,³, 段秋慧², 李大力²(), 陈金联³()

^1. 安徽理工大学医学院,淮南 232000
^2. 华东师范大学生命科学院,上海市调控生物学重点实验室,上海 200241
^3. 上海交通大学附属第六人民医院南院消化内科,上海 201499

收稿日期:2020-11-26 修回日期:2020-12-22 出版日期:2021-01-20 发布日期:2021-01-04
通讯作者: 李大力,陈金联 E-mail:dlli@bio.ecnu.edu.cn;wqq_021002@163.com
作者简介:王芯悦,在读硕士研究生,专业方向：消化内科学。E-mail: 1294008801@qq.com
基金资助:
上海市自然科学基金项目资助编号(16ZR1429100)

Effect of Uhrf1 on intestinal development

Xinyue Wang^1,³, Liang Li^2,³, Qiuhui Duan², Dali Li²(), Jinlian Chen³()

^1. School of Medicine, Anhui University of Science and Technology, Huainan 232000, China
^2. School of Life Sciences, East China Normal University, Shanghai Key Laboratory of Regulatory Biology, Shanghai 200241, China
^3. Department of Gastroenterology, South Campus of Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 201499, China

Received:2020-11-26 Revised:2020-12-22 Online:2021-01-20 Published:2021-01-04
Contact: Li Dali,Chen Jinlian E-mail:dlli@bio.ecnu.edu.cn;wqq_021002@163.com
Supported by:
Supported by the National Natural Science Foundation of Shanghai No(16ZR1429100)

摘要/Abstract

摘要：

作为一种常见的表观遗传修饰类型,DNA甲基化对哺乳动物发育起着重要作用。Uhrf1作为重要的表观遗传调控因子,在DNA合成过程中可结合半甲基化的DNA同时招募DNA甲基转移酶1参与DNA甲基化的维持,保证遗传信息在细胞分裂前后的稳定传递。目前关于Uhrf1介导的DNA甲基化是否影响肠上皮发育过程尚不清楚。为探索Uhrf1在肠上皮发育中的作用,本研究成功构建了肠上皮特异性敲除Uhrf1的小鼠模型,利用HE染色对肠上皮组织形态学观察发现,与正常小鼠相比,敲除Uhrf1的小鼠肠上皮发育异常,主要表现为绒毛变短,数量减少,隐窝萎缩;通过表型分析发现,在小鼠肠上皮中特异性敲除Uhrf1后,细胞增殖明显受到抑制、凋亡细胞增加、细胞分化异常,同时肠干细胞相关基因表达降低。进一步对可能的分子机制进行初步探索发现Uhrf1缺失后 DNA甲基化水平大幅下降,诱发DNA损伤。本研究结果表明Uhrf1介导的DNA甲基化对肠上皮的正常发育成熟具有重要作用,有望丰富Uhrf1介导的DNA甲基化在体内的生物学功能,并为进一步明确Uhrf1介导的表观遗传调控机制提供实验依据。

关键词: DNA甲基化, Uhrf1, 肠上皮发育

Abstract:

As a best-characterized epigenetic modification, DNA methylation plays an important role in mammalian development. Uhrf1 is a critical epigenetic regulator that can bind to hemimethylated DNA and recruit DNA methyltransferase 1 to maintain DNA methylation. So far, the role of Uhrf1-mediated DNA methylation in intestinal development is still unknown. In order to investigate the impact of Uhrf1 deletion in intestinal development, we have successfully constructed the epithelial-specific Uhrf1 knockout mouse model. After Uhrf1 ablation, we found the mutant mice exhibited abnormal epithlial structure with less and shorter villi and shrinked crypts compared with wild type mice via hematoxylin-eosin staining. Further analysis showed that Uhrf1 deletion in the intestinal epithelium significantly decreased the cell proliferation and induced cell apoptosis. In addition, Uhrf1 deletion inhibited the normal epithelial differentiation and the expression of intestinal stem cell marker genes. Preliminary mechanism study revealed that loss of Uhrf1 caused global DNA hypomethylation which induced DNA damage in crypt cells. Taken together, our data suggested that DNA methylation mediated by Uhrf1 is vital for the normal intestinal development. Our results enriched the in vivo role of Uhrf1 and laid the foundation for further epigenetic regulatory mechanism exploration.

Key words: DNA methylation, Uhrf1, intestinal development

王芯悦, 李亮, 段秋慧, 李大力, 陈金联. Uhrf1对肠上皮发育的影响[J]. 遗传, 2021, 43(1): 84-93.

Xinyue Wang, Liang Li, Qiuhui Duan, Dali Li, Jinlian Chen. Effect of Uhrf1 on intestinal development[J]. Hereditas(Beijing), 2021, 43(1): 84-93.

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基因	引物序列(5′→3′)
Lgr5	F: CAGTGTTGTGCATTTGGGGG
Lgr5	R: CAAGGTCCCGCTCATCTTGA
Sox9	F: CACAAGAAAGACCACCCCGA
Sox9	R: GGACCCTGAGATTGCCCAGA
Ascl2	F: CGTGAAGCTGGTGAACTTGG
Ascl2	R: GGATGTACTCCACGGCTGAG
Olfm4	F: TCTTGGGCAGAAGGTGGGACT
Olfm4	R: GGACCGTCAGGTTCAGGAGC
Uhrf1	F: ACGGTGCCTACTCATTGGTC
Uhrf1	R: GCTTCTGGTCAGAGGACTGG
β-actin	F: CAGCCTTCCTTCTTGGGTAT
β-actin	R: TGATCTTGATCTTCATGGTGC

Uhrf1对肠上皮发育的影响

Effect of Uhrf1 on intestinal development

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