RhoA promotes epidermal stem cell proliferation via PKN1-cyclin D1 signaling

Wang, Fan; Zhan, Rong; Chen, Liang; Dai, Xin; Wang, Wenping; Guo, Rui; Li, Xiaoge; Li, Zhe; Wang, Liang; Huang, Shupeng; Shen, Jie; Liu, Shirong; Cao, Chuan

doi:10.1371/journal.pone.0172613

Cited by 18 publications

(20 citation statements)

References 41 publications

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“…2A). Wang fan et al found that cyclin D1 was signi cantly expressed during the proliferation of human EpSCs [29]. We previously reported that nitric oxide induces FoxG1 expression in human EpSCs [32].…”

Section: Discussionmentioning

confidence: 91%

“…To analyze the effect of GDF-5 on EpSCs proliferation in vivo, BrdU-labelled EpSCs and mouse model of burn injury were established as our previously described [29]. BrdU + and PCNA + EpSCs were presented by immunochemistry in the regenerated epidermis, and the double positive EpSCs were counted in the different re-epithelialization area.…”

Section: The Effect Of Gdf-5 On Mouse Epscs Proliferation Via Foxg1/cmentioning

confidence: 99%

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GDF-5 promotes Epidermal Stem Cells proliferation via Foxg1-cyclin D1 signaling

Zhao

Bian²,

Wang³

et al. 2020

Preprint

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Objective: Epidermal stem cells (EpSCs) can self-renew and are responsible for the long-term maintenance of the skin, it also plays a critical role in wound re-epithelization, but the mechanism underlying EpSCs proliferation is unclear. Here, we studied the effects of GDF-5 on mouse EpSCs proliferation mechanism in wound healing. Methods: Firstly, the effects of GDF-5 on EpSCs proliferation was tested by using CCK8 reagent and PCNA expression was analysed by western blotting. Secondly, we screened genes that promote EpSCs proliferation in the FOX family and cyclin by qPCR, and further analysed the protein expression level of the selected genes by Western blotting. Thirdly, siRNA plasmids and pAdEasy adenovirus were transfected or infected, respectively, into mouse EpSCs to detect the effect of target genes on GDF-5 induced cell proliferation. Furthermore, a deep partial thickness burn mouse model was used in which GDF-5 induced EpSCs proliferation was detected by immunohistochemical. Finally, the relationships target genes were analysed by qPCR, immunoblotting and dual luciferase reporter gene detection. Results: We discovered that 100 ng/ml recombinant mouse GDF-5 was the optimal concentration for promoting mouse EpSCs proliferation. Through preliminary screening by qPCR, we found that Foxg1 and cyclin D1 could be the downstream molecules of GDF-5, and the results were confirmed by Western blotting. And the effect of GDF-5 on mouse EpSCs proliferation was regulated by Foxg1/cyclin D1 in vitro and in vivo. Besides, GDF-5 induced transcription of cyclin D1 was regulated by Foxg1-mediated cyclin D1 promoter activity.Conclusion: This paper showed that GDF-5 promotes mouse EpSCs proliferation via Foxg1-cyclin D1 signal pathway. It is suggested that GDF-5 may be a new approach to yield EpSCs for wound healing.

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Section: Discussionmentioning

confidence: 91%

Section: The Effect Of Gdf-5 On Mouse Epscs Proliferation Via Foxg1/cmentioning

confidence: 99%

GDF-5 promotes Epidermal Stem Cells proliferation via Foxg1-cyclin D1 signaling

Zhao

Bian²,

Wang³

et al. 2020

Preprint

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“…Besides, the Wnt/cyclin D1 pathway has a dedifferentiating effect for differentiated epidermal cells [ 28 ]. In our previous research, we found that cyclin D1 is an important downstream signaling molecule in the proliferation of EpSCs [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

GDF-5 promotes epidermal stem cells proliferation via Foxg1-cyclin D1 signaling

et al. 2021

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Objective Epidermal stem cells (EpSCs) can self-renew, which are responsible for the long-term maintenance of the skin, and it also plays a critical role in wound re-epithelization, but the mechanism underlying EpSCs proliferation is unclear. GDF-5, also known as BMP-14, is a member of the BMP family and can be used as a self-renewal supporter. Here, we studied the effects of GDF-5 on mouse EpSCs proliferation mechanism in wound healing. Methods Firstly, the effects of GDF-5 on EpSCs proliferation was tested by using CCK8 reagent and PCNA expression was analyzed by Western blotting. Secondly, we screened genes that promote EpSCs proliferation in the FOX and cyclin family by qPCR, and then the protein expression level of the selected genes was further analyzed by Western blotting. Thirdly, siRNA plasmids and pAdEasy adenovirus were transfected or infected, respectively, into mouse EpSCs to detect the effect of target genes on GDF-5-induced cell proliferation. Furthermore, we injected GDF-5 to a deep partial thickness burn mouse model for finding out whether EpSCs proliferation can be detected by immunohistochemical. Finally, the relevant target genes were analyzed by qPCR, immunoblotting, and dual-luciferase reporter gene detection. Results We discovered that 100 ng/ml recombinant mouse GDF-5 was the optimal concentration for promoting mouse EpSCs proliferation. Through preliminary screened by qPCR, we found that Foxg1 and cyclin D1 could be the downstream molecules of GDF-5, and the results were confirmed by Western blotting. And the effect of GDF-5 on mouse EpSCs proliferation was adjusted by Foxg1/cyclin D1 in vitro and in vivo. Besides, GDF-5-induced transcription of cyclin D1 was regulated by Foxg1-mediated cyclin D1 promoter activity. Conclusion This paper showed that GDF-5 promotes mouse EpSCs proliferation via Foxg1-cyclin D1 signal pathway. It is suggested that GDF-5 may be a new approach to make EpSCs proliferation which can be used in wound healing.

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“…The CyTOF allows for a wide staining panel, capturing several cell groups in detail and providing a systemic view of differentiation. Whereas IdU injection in mice has been used in microscopy imaging, for neogenesis (14), cell cycle analysis (15), and proliferation (16), its use in single-cell mass cytometry remains limited. IdU has been used in vitro on CyTOF measurements (17) to study cell cycle and to immunologically profile proliferating versus non-proliferating cells in the mouse bone marrow (18), however, it remains an untapped resource to study differentiation.…”

Section: Introductionmentioning

confidence: 99%

Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse‐Chase, Mass Cytometry, and Mathematical Modeling

2019

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We present a new method to directly quantify the dynamics of differentiation of multiple cellular subsets in unperturbed mice. We combine a pulse-chase protocol of IdU injections with subsequent analysis by mass cytometry (CyTOF), and mathematical modeling of the IdU dynamics. Measurements by CyTOF allow for a wide range of cells to be analyzed at once, due to the availability of a large staining panel without the complication of fluorescence spill-over. These are also compatible with direct detection of integrated iodine signal, with minimal impact on immunophenotyping based on surface markers. Mathematical modeling beyond a binary classification of surface marker abundance allows for a continuum of cellular states as the cells transition from one state to another. Thus, we present a complete and robust method for directly quantifying differentiation at the systemic level, allowing for system-wide comparisons between different mouse strains and/or experimental conditions.

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RhoA promotes epidermal stem cell proliferation via PKN1-cyclin D1 signaling

Cited by 18 publications

References 41 publications

GDF-5 promotes Epidermal Stem Cells proliferation via Foxg1-cyclin D1 signaling

GDF-5 promotes Epidermal Stem Cells proliferation via Foxg1-cyclin D1 signaling

GDF-5 promotes epidermal stem cells proliferation via Foxg1-cyclin D1 signaling

Quantifying the Dynamics of Hematopoiesis by In Vivo IdU Pulse‐Chase, Mass Cytometry, and Mathematical Modeling

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