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Title: Hedgehog signaling contributes to basic fibroblast growth factor-regulated fibroblast migration

Abstract

Fibroblast migration is a central process in skin wound healing, which requires the coordination of several types of growth factors. bFGF, a well-known fibroblast growth factor (FGF), is able to accelerate fibroblast migration; however, the underlying mechanism of bFGF regulation fibroblast migration remains unclear. Through the RNA-seq analysis, we had identified that the hedgehog (Hh) canonical pathway genes including Smoothened (Smo) and Gli1, were regulated by bFGF. Further analysis revealed that activation of the Hh pathway via up-regulation of Smo promoted fibroblast migration, invasion, and skin wound healing, but which significantly reduced by GANT61, a selective antagonist of Gli1/Gli2. Western blot analyses and siRNA transfection assays demonstrated that Smo acted upstream of phosphoinositide 3-kinase (PI3K)-c-Jun N-terminal kinase (JNK)-β-catenin to promote cell migration. Moreover, RNA-seq and qRT-PCR analyses revealed that Hh pathway genes including Smo and Gli1 were under control of β-catenin, suggesting that β-catenin turn feedback activates Hh signaling. Taken together, our analyses identified a new bFGF-regulating mechanism by which Hh signaling regulates human fibroblast migration, and the data presented here opens a new avenue for the wound healing therapy. - Highlights: • bFGF regulates Hedgehog (Hh) signaling in fibroblasts. • The Smo and Gli two master regulators of Hhmore » signaling positively regulate fibroblast migration. • Smo facilitates β-catenin nuclear translocation via activation PI3K/JNK/GSK3β. • β-catenin positively regulates fibroblast cell migration and the expression of Hh signaling genes including Smo and Gli.« less

Authors:
 [1];  [1];  [2]; ; ; ; ;  [1];  [3];  [4];  [1];  [1];  [1]
  1. School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang (China)
  2. (China)
  3. Wenzhou People's Hospital, Wenzhou, Zhejiang (China)
  4. Ningbo First Hospital, Ningbo, Zhejiang (China)
Publication Date:
OSTI Identifier:
22649858
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 355; Journal Issue: 2; Other Information: Copyright (c) 2017 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; FIBROBLASTS; GENES; GROWTH FACTORS; HEALING; PHOSPHOTRANSFERASES; POLYMERASE CHAIN REACTION; RNA; SKIN; THERAPY; TRANSLOCATION; WOUNDS

Citation Formats

Zhu, Zhong Xin, Sun, Cong Cong, Wenzhou People's Hospital, Wenzhou, Zhejiang, Ting Zhu, Yu, Wang, Ying, Wang, Tao, Chi, Li Sha, Cai, Wan Hui, Zheng, Jia Yong, Zhou, Xuan, Cong, Wei Tao, Li, Xiao Kun, E-mail: proflxk@163.com, and Jin, Li Tai, E-mail: jin_litai@126.com. Hedgehog signaling contributes to basic fibroblast growth factor-regulated fibroblast migration. United States: N. p., 2017. Web. doi:10.1016/J.YEXCR.2017.03.054.
Zhu, Zhong Xin, Sun, Cong Cong, Wenzhou People's Hospital, Wenzhou, Zhejiang, Ting Zhu, Yu, Wang, Ying, Wang, Tao, Chi, Li Sha, Cai, Wan Hui, Zheng, Jia Yong, Zhou, Xuan, Cong, Wei Tao, Li, Xiao Kun, E-mail: proflxk@163.com, & Jin, Li Tai, E-mail: jin_litai@126.com. Hedgehog signaling contributes to basic fibroblast growth factor-regulated fibroblast migration. United States. doi:10.1016/J.YEXCR.2017.03.054.
Zhu, Zhong Xin, Sun, Cong Cong, Wenzhou People's Hospital, Wenzhou, Zhejiang, Ting Zhu, Yu, Wang, Ying, Wang, Tao, Chi, Li Sha, Cai, Wan Hui, Zheng, Jia Yong, Zhou, Xuan, Cong, Wei Tao, Li, Xiao Kun, E-mail: proflxk@163.com, and Jin, Li Tai, E-mail: jin_litai@126.com. 2017. "Hedgehog signaling contributes to basic fibroblast growth factor-regulated fibroblast migration". United States. doi:10.1016/J.YEXCR.2017.03.054.
@article{osti_22649858,
title = {Hedgehog signaling contributes to basic fibroblast growth factor-regulated fibroblast migration},
author = {Zhu, Zhong Xin and Sun, Cong Cong and Wenzhou People's Hospital, Wenzhou, Zhejiang and Ting Zhu, Yu and Wang, Ying and Wang, Tao and Chi, Li Sha and Cai, Wan Hui and Zheng, Jia Yong and Zhou, Xuan and Cong, Wei Tao and Li, Xiao Kun, E-mail: proflxk@163.com and Jin, Li Tai, E-mail: jin_litai@126.com},
abstractNote = {Fibroblast migration is a central process in skin wound healing, which requires the coordination of several types of growth factors. bFGF, a well-known fibroblast growth factor (FGF), is able to accelerate fibroblast migration; however, the underlying mechanism of bFGF regulation fibroblast migration remains unclear. Through the RNA-seq analysis, we had identified that the hedgehog (Hh) canonical pathway genes including Smoothened (Smo) and Gli1, were regulated by bFGF. Further analysis revealed that activation of the Hh pathway via up-regulation of Smo promoted fibroblast migration, invasion, and skin wound healing, but which significantly reduced by GANT61, a selective antagonist of Gli1/Gli2. Western blot analyses and siRNA transfection assays demonstrated that Smo acted upstream of phosphoinositide 3-kinase (PI3K)-c-Jun N-terminal kinase (JNK)-β-catenin to promote cell migration. Moreover, RNA-seq and qRT-PCR analyses revealed that Hh pathway genes including Smo and Gli1 were under control of β-catenin, suggesting that β-catenin turn feedback activates Hh signaling. Taken together, our analyses identified a new bFGF-regulating mechanism by which Hh signaling regulates human fibroblast migration, and the data presented here opens a new avenue for the wound healing therapy. - Highlights: • bFGF regulates Hedgehog (Hh) signaling in fibroblasts. • The Smo and Gli two master regulators of Hh signaling positively regulate fibroblast migration. • Smo facilitates β-catenin nuclear translocation via activation PI3K/JNK/GSK3β. • β-catenin positively regulates fibroblast cell migration and the expression of Hh signaling genes including Smo and Gli.},
doi = {10.1016/J.YEXCR.2017.03.054},
journal = {Experimental Cell Research},
number = 2,
volume = 355,
place = {United States},
year = 2017,
month = 6
}
  • The growth autonomy of human tumor cells is considered due to the endogenous production of growth factors. Transcriptional expression of candidates for autocrine stimulatory factors such as basic fibroblast growth factor (FGF), acidic FGF, and transforming growth factor type {beta} were determined in human brain tumors. Basic FGF was expressed abundantly in 17 of 18 gliomas, 20 of 22 meningiomas, and 0 of 5 metastatic brain tumors. The level of mRNA expression of acidic FGF in gliomas was significant. In contrast, transforming growth factor type {beta}1 was expressed in all the samples investigated. The mRNA for basic FGF and itsmore » peptide were localized in tumor cells in vivo by in situ hybridization and immunohistochemistry, showing that basic FGF is actually produced in tumor cells. The results suggest that tumor-derived basic FGF is involved in the progression of gliomas and meningiomas in vivo, whereas acidic FGF is expressed in a tumor origin-specific manner, suggesting that acidic FGF works in tandem with basic FGF in glioma tumorigenesis.« less
  • In contrast to the well-known cytotoxic effects of tumor necrosis factor a (TNF) in many mammary cancer cells, we have found that TNF stimulates the proliferation and motility of human mammary epithelial cells (HMEC). Since the response of HMEC to TNF is similar to effects mediated by epidermal growth factor receptor (EGFR) activation, we explored the potential role of cross-talk through the EGFR signaling pathways in mediating cellular responses to TNF. Using a microarray enzyme-linked immunoassay, we found that exposure to TNF stimulated the dose-dependent shedding of the EGFR ligand transforming growth factor a (TGFa). Both proliferation and motility ofmore » HMEC induced by TNF was prevented either by inhibiting membrane protein shedding with a metalloprotease inhibitor, by blocking EGFR kinase activity, or by limiting ligand-receptor interactions with an antagonistic anti-EGFR antibody. EGFR activity was also necessary for TNF-induced release of MMP-9, a matrix metalloprotease thought to be an essential regulator of mammary cell migration. The cellular response to TNF was associated with a biphasic temporal pattern of extracellular signal-regulated kinase (ERK) phosphorylation, which was EGFR-dependent and modulated by inhibition of metalloprotease-mediated shedding. Significantly, the late phase of ERK phosphorylation, detectable within 4 hours after exposure, was blocked by the metalloprotease inhibitor batimastat, indicating that autocrine signaling through ligand shedding was responsible for this secondary wave of ERK activity. Our results indicate a novel and important role for metalloprotease activation and EGFR transmodulation in mediating the cellular response to TNF.« less
  • Keratinocyte growth factor (KGF/FGF7) and fibroblast growth factor 10 (FGF10/KGF2) regulate keratinocyte proliferation and differentiation by binding to the tyrosine kinase KGF receptor (KGFR). KGF induces keratinocyte motility and cytoskeletal rearrangement, whereas a direct role of FGF10 on keratinocyte migration is not clearly established. Here we analyzed the motogenic activity of FGF10 and KGF on human keratinocytes. Migration assays and immunofluorescence of actin cytoskeleton revealed that FGF10 is less efficient than KGF in promoting migration and exerts a delayed effect in inducing lamellipodia and ruffles formation. Both growth factors promoted phosphorylation and subsequent membrane translocation of cortactin, an F-actin bindingmore » protein involved in cell migration; however, FGF10-induced cortactin phosphorylation was reduced, more transient and delayed with respect to that promoted by KGF. Cortactin phosphorylation induced by both growth factors was Src-dependent, while its membrane translocation and cell migration were blocked by either Src and PI3K inhibitors, suggesting that both pathways are involved in KGF- and FGF10-dependent motility. Furthermore, siRNA-mediated downregulation of cortactin inhibited KGF- and FGF10-induced migration. These results indicate that cortactin is involved in keratinocyte migration promoted by both KGF and FGF10.« less
  • To determine if fibroblasts could be a source of fibroblast growth factor (FGF) in tissue, cells were initiated in culture from newborn human foreskin. Fibroblast cell lysates promoted radiolabeled thymidine uptake by cultured quiescent fibroblasts. Seventy-nine percent of the growth-promoting activity of lysates was recovered from heparin-Sepharose. The heparin-binding growth factor reacted on immunoblots with antiserum to human placenta-derived basic FGF and competed with iodinated basic FGF for binding to antiserum to (1-24)bFGF synthetic peptide. To confirm that fibroblasts were the source of the growth factor, cell lysates were prepared from cells incubated with radiolabeled methionine. Heparin affinity purified materialmore » was immunoprecipitated with basic FGF antiserum and electrophoresed. Radiolabeled material was detected on gel autoradiographs in the same molecular weight region as authentic iodinated basic FGF. The findings are consistant with the notion that cultured fibroblasts express basic FGF. As these cells also respond to the mitogen, it is possible that the regulation of their growth is under autocrine control. Fibroblasts may be an important source of the growth factor in tissue.« less
  • No abstract prepared.