Ten-eleven translocation 1 mediating DNA demethylation regulates the proliferation of chicken primordial germ cells through the activation of Wnt4/β-catenin signaling pathway
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Objective: The objective of this study was to investigate the regulation relationship of Teneleven translocation 1 (<i>Tet1</i>) in DNA demethylation and the proliferation of primordial germ cells (PGCs) in chickens.Methods: siRNA targeting <i>Tet1</i> was used to transiently knockdown the expression of <i>Tet1</i> in chicken PGCs, and the genomic DNA methylation status was measured. The proliferation of chicken PGCs was detected by flow cytometry analysis and cell counting kit-8 assay when activation or inhibition of Wnt4/β-catenin signaling pathway. And the level of DNA methylation and hisotne methylation was also tested.Results: Results revealed that knockdown of <i>Tet1</i> inhibited the proliferation of chicken PGCs and downregulated the mRNA expression of <i>Cyclin D1</i> and cyclin-dependent kinase 6 (<i>CDK6</i>), as well as pluripotency-associated genes (<i>Nanog, PouV</i>, and <i>Sox2</i>). Flow cytometry analysis confirmed that the population of PGCs in <i>Tet1</i> knockdown group displayed a significant decrease in the proportion of S and G2 phase cells, which meant that there were less PGCs entered the mitosis process than that of control. Furthermore, <i>Tet1</i> knockdown delayed the entrance to G1/S phase and this inhibition was rescued by treated with BIO. Consistent with these findings, Wnt/β-catenin signaling was inactivated in <i>Tet1</i> knockdown PGCs, leading to aberrant proliferation. Further analysis showed that the methylation of the whole genome increased significantly after <i>Tet1</i> downregulation, while hydroxymethylation obviously declined. Meanwhile, the level of H3K27me3 was upregulated and H3K9me2 was downregulated in <i>Tet1</i> knockdown PGCs, which was achieved by regulating Wnt/β-catenin signaling pathway.Conclusion: These results suggested that the self-renewal of chicken PGCs and the maintenance of their characteristics were regulated by <i>Tet1</i> mediating DNA demethylation through the activation of Wnt4/β-catenin signaling pathway.
Abstract
Objective: The objective of this study was to investigate the regulation relationship of Teneleven translocation 1 (Tet1) in DNA demethylation and the proliferation of primordial germ cells (PGCs) in chickens. Methods: siRNA targeting Tet1 was used to transiently knockdown the expression of Tet1 in chicken PGCs, and the genomic DNA methylation status was measured. The proliferation of chicken PGCs was detected by flow cytometry analysis and cell counting kit-8 assay when activation or inhibition of Wnt4/β-catenin signaling pathway. And the level of DNA methylation and hisotne methylation was also tested. Results: Results revealed that knockdown of Tet1 inhibited the proliferation of chicken PGCs and downregulated the mRNA expression of Cyclin D1 and cyclin-dependent kinase 6 (CDK6), as well as pluripotency-associated genes (Nanog, PouV, and Sox2). Flow cytometry analysis confirmed that the population of PGCs in Tet1 knockdown group displayed a significant decrease in the proportion of S and G2 phase cells, which meant that there were less PGCs entered the mitosis process than that of control. Furthermore, Tet1 knockdown delayed the entrance to G1/S phase and this inhibition was rescued by treated with BIO. Consistent with these findings, Wnt/β-catenin signaling was inactivated in Tet1 knockdown PGCs, leading to aberrant proliferation. Further analysis showed that the methylation of the whole genome increased significantly after Tet1 downregulation, while hydroxymethylation obviously declined. Meanwhile, the level of H3K27me3 was upregulated and H3K9me2 was downregulated in Tet1 knockdown PGCs, which was achieved by regulating Wnt/β-catenin signaling pathway. Conclusion: These results suggested that the self-renewal of chicken PGCs and the maintenance of their characteristics were regulated by Tet1 mediating DNA demethylation through the activation of Wnt4/β-catenin signaling pathway.
참고문헌 (35)
- Ramakrishna NB, "Epigenetic regulation during primordial germ cell development and differentiation", Sex Dev, 15, pp.411~431 (2021)[DOI]
- Hancock GV, "Mammalian primordial germ cell specification", Development, 148, pp.dev189217~ (2021)[DOI]
- Wang XX, "The role of DNA methylation reprogramming during sex determination and transition in zebrafish", Genomics Proteomics Bioinformatics, 19, pp.48~63 (2021)[DOI]
- Wang X, "The dynamics of DNA methylation during epigenetic reprogramming of primordial germ cells in medaka(Oryzias latipes)", Epigenetics, 15, pp.483~498 (2020)[DOI]
- Yu ML, "Effects of teneleven translocation 1(Tet1)on DNA methylation and gene expression in chicken primordial germ cells", Reprod Fertil Dev, 31, pp.509~520 (2019)[DOI]
- Gallego-Bartolome J, "Targeted DNA demethylation of the Arabidopsis genome using the human TET1 catalytic domain", Proc Natl Acad Sci USA, 115, pp.E2125~E2134 (2018)[DOI]
- Zuo QS, "Identification of key events and regulatory networks in the formation process of primordial germ cell based on proteomics", J Cell Physiol, 238, pp.610~630 (2023)[DOI]
- Kohli RM, "TET enzymes, TDG and the dynamics of DNA demethylation", Nature, 502, pp.472~479 (2013)[DOI]
- Khoueiry R, "Lineage-specific functions of TET1 in the postimplantation mouse embryo", Nat Genet, 49, pp.1061~1072 (2017)[DOI]
- Gavin DP, "Active DNA demethylation in post-mitotic neurons : a reason for optimism", Neuropharma"cology, 75, pp.233~245 (2013)[DOI]
- Ito S, "Role of Tet proteins in 5mC to 5hmC conversion, ES-cell self-renewal and inner cell mass specification", Nature, 466, pp.1129~1133 (2010)[DOI]
- Miyoshi N, "Erasure of DNA methylation, genomic imprints, and epimutations in a primordial germ-cell model derived from mouse pluripotent stem cells", Proc Natl Acad Sci USA, 113, pp.9545~9550 (2016)[DOI]
- Ohinata Y, "A signaling principle for the specification of the germ cell lineage in mice", Cell, 137, pp.571~584 (2009)[DOI]
- Kai M, "TET1 depletion induces aberrant CpG methylation in colorectal cancer cells", PLoS One, 11, pp.e0168281~ (2016)[DOI]
- Jiang X, "DACT3 is an epigenetic regulator of Wnt", beta-catenin signaling in colorectal cancer and is a therapeutic target of histone modifications, 2008, pp.13, 529~541
- Waaler J, "Novel synthetic antagonists of canonical Wnt signaling inhibit colorectal cancer cell growth", Cancer Res, 71, pp.197~205 (2011)[DOI]
- Zhang B, "Knockdown of Homeobox B5(HOXB5)inhibits cell proliferation, migration, and invasion in non-small cell lung cancer cells through inactivation of the Wnt", beta-catenin pathway, 2018, pp.26, 37~44
- Zhang C, "H3K4me2 promotes the activation of lncCPSET1 by Jun in the chicken PGC formation", Animals(Basel), 11, pp.1572~ (2021)[DOI]
- D'Orazio FM, "Germ cell differentiation requires Tdrd7-dependent chromatin and transcriptome reprogramming marked by germ plasm relocalization", Dev Cell, 56, pp.641~656 (2021)[DOI]
- Kress C, "Chicken embryonic stem cells and primordial germ cells display different heterochromatic histone marks than their mammalian counterparts", Epigenetics Chromatin, 9, pp.5~ (2016)[DOI]
- Mallol A, "PRDM14 controls X-chromosomal and global epigenetic reprogramming of H3K27me3 in migrating mouse primordial germ cells", Epigenetics Chromatin, 12, pp.38~ (2019)[DOI]
- Hajkova P, "Genome-wide reprogramming in the mouse germ line entails the base excision repair pathway", Science, 329, pp.78~82 (2010)[DOI]
- Yu ML, "Retinoic acid promotes proliferation of chicken primordial germ cells via activation of PI3K", Akt-mediated NF-kappaB signalling cascade, 2012, pp.36, 705~712
- Yamaguchi S, "Role of Tet1 in erasure of genomic imprinting", Nature, 504, pp.460~464 (2013)[DOI]
- Matsui Y, "A current view of the epigenome in mouse primordial germ cells", Mol Reprod Dev, 81, pp.160~170 (2014)[DOI]
- Tahiliani M, "Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1", Science, 324, pp.930~935 (2009)[DOI]
- Lian CG, "Loss of 5-hydroxymethylcytosine is an epigenetic hallmark of melanoma", Cell, 150, pp.1135~1146 (2012)[DOI]
- Hackett JA, "Synergistic mechanisms of DNA demethylation during transition to ground-state pluripotency", Stem Cell Rep, 1, pp.518~531 (2013)[DOI]
- Piccolo FM, "Different roles for Tet1 and Tet2 proteins in reprogramming-mediated erasure of imprints induced by EGC fusion", Mol Cell, 49, pp.1176~ (2013)[DOI]
- Guibert S, "Global profiling of DNA methylation erasure in mouse primordial germ cells", Genome Res, 22, pp.633~641 (2012)[DOI]
- Lee HC, "Wnt", β-catenin signaling pathway activation is required for proliferation of chicken primordial germ cells in vitro, 2016, pp.6, 34510~
- Zhang RR, "Tet1 regulates adult hippocampal neurogenesis and cognition", Cell Stem Cell, 13, pp.237~245 (2013)[DOI]
- Dai HQ, "TET-mediated DNA demethylation controls gastrulation by regulating Lefty-Nodal signalling", Nature, 538, pp.528~532 (2016)[DOI]
- Jin K, "CREPT and p15RS regulate cell proliferation and cycling in chicken DF-1 cells through the Wnt", beta-catenin pathway, 2018, pp.119, 1083~1092
- Zhang L, "H19knockdown suppresses proliferation and induces apoptosis by regulating miR-148b", WNT, J Biomed Sci, pp.2018, 25[DOI]