Circ0001470 Acts as a miR-140-3p Sponge to Facilitate the Progression of Embryonic Development through Regulating PTGFR Expression

Long Zhang; Changfan Zhou; Xiaoyu Jiang; Shuntao Huang; Yiheng Li; Tao Su; Guowei Wang; You Zhou; Min Liu; Dequan Xu

doi:10.3390/cells11111746

Circ0001470 Acts as a miR-140-3p Sponge to Facilitate the Progression of Embryonic Development through Regulating PTGFR Expression

Cells. 2022 May 25;11(11):1746. doi: 10.3390/cells11111746.

Authors

Long Zhang^{1

2

3}, Changfan Zhou^{1

2

3}, Xiaoyu Jiang^{1

2

3}, Shuntao Huang^{1

2

3}, Yiheng Li¹, Tao Su^{1

2

3}, Guowei Wang^{1

2

3}, You Zhou¹, Min Liu^{1

4}, Dequan Xu^{1

2

3}

Affiliations

¹ Colleges of Animal Science & Technology, Huazhong Agricultural University, Wuhan 430070, China.
² Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, China.
³ Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China.
⁴ College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.

Abstract

Embryonic implantation and development are vital in early pregnancy and assisted reproduction. Circular RNAs (circRNAs) are involved in the two physiological processes and thus regulate animal reproduction. However, their specific regulatory functions and mechanisms remain unclear. Here, a novel circ0001470, originating from the porcine GRN gene, differentially expressed on day 18 versus day 32 of gestation in Meishan and Yorkshire pigs was screened. The circularization characteristic of circ0001470 was identified based on divergent primer amplification, Sanger sequencing, RNase digestion, and RNA nuclear-cytoplasmic fractionation. Functionally, circ0001470 can promote cell proliferation and cycle progression of endometrial epithelial cells (EECs) and also inhibit apoptosis of EECs using CCK-8 assays and flow cytometry analyses. Mechanistically, bioinformatics database prediction, luciferase screening, RNA immunoprecipitation (RIP), RNA-pull down, and FISH co-localization experiments revealed that the circ0001470 acted as a competing endogenous RNA (ceRNA) through sponging miR-140-3p to regulate downstream PTGFR expression. Moreover, in vivo assays revealed that mmu_circGRN promoted embryonic development by affecting the expression of PTGFR, which can activate the MAPK reproduction pathway and facilitate pregnancy maintenance. This study enriched our understanding of circRNAs in embryo implantation and development by deciding the fate of EECs.

Keywords: circ0001470; embryonic development; endometrial epithelial cells; miR-140-3p; proliferation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
Cell Proliferation / genetics
Embryonic Development / genetics
MicroRNAs* / genetics
MicroRNAs* / metabolism
RNA, Circular* / genetics
Receptors, Prostaglandin / metabolism
Swine

Substances

MicroRNAs
RNA, Circular
Receptors, Prostaglandin
prostaglandin F2alpha receptor

Grants and funding

This research was funded by National Key Research and Development Program of China (2021YFD1301201), Fundamental Research Funds for the Central Universities (2662020DKPY003), Agricultural Science Innovation Foundation of Hubei Province (2019-620-000-001-30), State Key Development Program for Basic Research of China (2014CB138504), National Science and Technology Major Project of China (2016ZX08006002), Animal Husbandry Major Technology Collaborative Extension Project of Hubei Province, Research Project of Huazhong Agricultural University, China.