title: Epidermal Growth Factor Induces Proliferation of Hair Follicle-Derived Mesenchymal Stem Cells Through Epidermal Growth Factor Receptor-Mediated Activation of ERK and AKT Signaling Pathways Associated with Upregulation of Cyclin D1 and Downregulation of p16.
abstract: The maintenance of highly proliferative capacity and full differentiation potential is a necessary step in the initiation of stem cell-based regenerative medicine. Our recent study showed that epidermal growth factor (EGF) significantly enhanced hair follicle-derived mesenchymal stem cell (HF-MSC) proliferation while maintaining the multilineage differentiation potentials. However, the underlying mechanism remains unclear. Herein, we investigated the role of EGF in HF-MSC proliferation. HF-MSCs were isolated and cultured with or without EGF. Immunofluorescence staining, flow cytometry, cytochemistry, and western blotting were used to assess proliferation, cell signaling pathways related to the EGF receptor (EGFR), and cell cycle progression. HF-MSCs exhibited surface markers of mesenchymal stem cells and displayed trilineage differentiation potentials toward adipocytes, chondrocytes, and osteoblasts. EGF significantly increased HF-MSC proliferation as well as EGFR, ERK1/2, and AKT phosphorylation (p-EGFR, p-ERK1/2, and p-AKT) in a time- and dose-dependent manner, but not STAT3 phosphorylation. EGFR inhibitor (AG1478), PI3K-AKT inhibitor (LY294002), ERK inhibitor (U0126), and STAT3 inhibitor (STA-21) significantly blocked EGF-induced HF-MSC proliferation. Moreover, AG1478, LY294002, and U0126 significantly decreased p-EGFR, p-AKT, and p-ERK1/2 expression. EGF shifted HF-MSCs at the G1 phase to the S and G2 phase. Concomitantly, cyclinD1, phosphorylated Rb, and E2F1expression increased, while that of p16 decreased. In conclusion, EGF induces HF-MSC proliferation through the EGFR/ERK and AKT pathways, but not through STAT-3. The G1/S transition was stimulated by upregulation of cyclinD1 and inhibition of p16 expression.
authors.affiliation: 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .|2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Tingting
authors.initials: T
authors.lastname: Bai
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .|3 Department of Ophthalmology, Second Hospital of Jilin University , Changchun, China .
authors.forename: Feilin
authors.initials: F
authors.lastname: Liu
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .|4 Department of Pediatrics, First Hospital of Jilin University , Changchun, China .
authors.forename: Fei
authors.initials: F
authors.lastname: Zou
authors.affiliation: 5 Department of Pathology, Jilin Medical College , Jilin, China .
authors.forename: Guifang
authors.initials: G
authors.lastname: Zhao
authors.affiliation: 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .|2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Yixu
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .|2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Jiahong
authors.initials: J
authors.lastname: Shi
authors.affiliation: 1 Department of Pathobiology, Key Laboratory of Ministry of Education, College of Basic Medicine, Jilin University , Changchun, China .|2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Deshun
authors.initials: D
authors.lastname: Hao
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Tong
authors.initials: T
authors.lastname: Zheng
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Yingyao
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Mingsheng
authors.initials: M
authors.lastname: Liu
authors.affiliation: 6 Department of Oncology, First People's Hospital of Lishu County , Lishu County, China .
authors.forename: Shilun
authors.initials: S
authors.lastname: Li
authors.affiliation: 7 Department of Thoracic Surgery, China-Japan Union Hospital of Jilin University , Changchun, China .
authors.forename: Liangchen
authors.initials: L
authors.lastname: Qi
authors.affiliation: 2 Department of Toxicology, School of Public Health, Jilin University , Changchun, China .
authors.forename: Jin Yu
authors.initials: JY
authors.lastname: Liu
mesh_terms: D000328:Adult; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D019938:Cyclin D1 / Q000378:metabolism*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D066246:ErbB Receptors / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005260:Female; D018859:Hair Follicle / Q000166:cytology*; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D008875:Middle Aged; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D019938:Cyclin D1; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases
keywords: EGFR; cell cycle; hair follicle; mesenchymal stem cells; proliferation