TGF-β-induced activation of conjunctival fibroblasts is modulated by FGF-2 and substratum stiffness

This study aimed to investigate the effects of substratum stiffness on the sensitivity of human conjunctival fibroblasts to transforming growth factor (TGF)-β, and to explore the molecular mechanism of action. Human conjunctival fibroblasts were cultured on collagen-coated plastic or silicone plates...

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Published in:PloS one 2020-11, Vol.15 (11), p.e0242626-e0242626
Main Authors: Matsumura, Tomoyo, Fujimoto, Tomokazu, Futakuchi, Akiko, Takihara, Yuji, Watanabe-Kitamura, Fumika, Takahashi, Eri, Inoue-Mochita, Miyuki, Tanihara, Hidenobu, Inoue, Toshihiro
Format: Article
Language:English
Subjects:
Actin
Actins - biosynthesis
Adaptor Proteins, Signal Transducing - metabolism
Antibodies
Biology and Life Sciences
Cell growth
Cell Line
Cell Proliferation
Cell Transdifferentiation
Collagen
Collagen (type I)
Collagen Type I - biosynthesis
Conjunctiva
Experiments
Fibroblast growth factor 2
Fibroblast Growth Factor 2 - metabolism
Fibroblasts
Fibronectin
Gene Expression Regulation
Glaucoma
Growth factors
Humans
Kinases
Laboratories
Life sciences
Medicine and Health Sciences
Muscles
Myofibroblasts - cytology
Myofibroblasts - metabolism
Physical Sciences
Plates
Proteins
R&D
Research & development
Silicone resins
Silicones
Smooth muscle
Stiffness
Surgery
Transcription
Transcription Factors - metabolism
Transforming Growth Factor beta2 - metabolism
Transforming Growth Factor beta2 - pharmacology
Wound healing
Yes-associated protein
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cited_by cdi_FETCH-LOGICAL-c526t-c3f528ae4aa81b1dee0eb49221b5b1cf1651197a06b409702b2d109a2ec3d7b63
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creator Matsumura, Tomoyo
Fujimoto, Tomokazu
Futakuchi, Akiko
Takihara, Yuji
Watanabe-Kitamura, Fumika
Takahashi, Eri
Inoue-Mochita, Miyuki
Tanihara, Hidenobu
Inoue, Toshihiro
description This study aimed to investigate the effects of substratum stiffness on the sensitivity of human conjunctival fibroblasts to transforming growth factor (TGF)-β, and to explore the molecular mechanism of action. Human conjunctival fibroblasts were cultured on collagen-coated plastic or silicone plates. The stiffness of the silicone plates was 0.2 or 64 kPa. Cells were treated by 2.5 ng/mL TGF-β2 with or without fibroblast growth factor (FGF)-2 (0-100 ng/mL) for 24 h or 48 h. The protein expression levels were determined by Western blot analysis. Cell proliferation was assessed using the WST-8 assay. FGF-2 suppressed the TGF-β-induced expression of α-smooth muscle actin (SMA) and collagen type I (Col I), but not fibronectin (FN). Both FGF-2 and TGF-β2 increased cell proliferation without an additive effect. The induction of α-SMA by TGF-β2 was decreased on the soft substratum, without any change in the expression level or subcellular location of Yes-associated protein/transcriptional coactivator with PDZ-binding motif (YAP/TAZ). FGF-2 suppressed TGF-β-induced α-SMA expression even on the soft substratum. FGF-2 treatment and a soft substratum suppressed TGF-β-induced transdifferentiation of conjunctival fibroblasts into myofibroblasts. FGF-2 attenuated the TGF-β-induced expression of α-SMA, even on a soft substratum.
doi_str_mv 10.1371/journal.pone.0242626
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Human conjunctival fibroblasts were cultured on collagen-coated plastic or silicone plates. The stiffness of the silicone plates was 0.2 or 64 kPa. Cells were treated by 2.5 ng/mL TGF-β2 with or without fibroblast growth factor (FGF)-2 (0-100 ng/mL) for 24 h or 48 h. The protein expression levels were determined by Western blot analysis. Cell proliferation was assessed using the WST-8 assay. FGF-2 suppressed the TGF-β-induced expression of α-smooth muscle actin (SMA) and collagen type I (Col I), but not fibronectin (FN). Both FGF-2 and TGF-β2 increased cell proliferation without an additive effect. The induction of α-SMA by TGF-β2 was decreased on the soft substratum, without any change in the expression level or subcellular location of Yes-associated protein/transcriptional coactivator with PDZ-binding motif (YAP/TAZ). FGF-2 suppressed TGF-β-induced α-SMA expression even on the soft substratum. FGF-2 treatment and a soft substratum suppressed TGF-β-induced transdifferentiation of conjunctival fibroblasts into myofibroblasts. FGF-2 attenuated the TGF-β-induced expression of α-SMA, even on a soft substratum.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>33206726</pmid><doi>10.1371/journal.pone.0242626</doi><orcidid>https://orcid.org/0000-0003-3769-7609</orcidid><oa>free_for_read</oa></addata></record>
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subjects Actin
Actins - biosynthesis
Adaptor Proteins, Signal Transducing - metabolism
Antibodies
Biology and Life Sciences
Cell growth
Cell Line
Cell Proliferation
Cell Transdifferentiation
Collagen
Collagen (type I)
Collagen Type I - biosynthesis
Conjunctiva
Experiments
Fibroblast growth factor 2
Fibroblast Growth Factor 2 - metabolism
Fibroblasts
Fibronectin
Gene Expression Regulation
Glaucoma
Growth factors
Humans
Kinases
Laboratories
Life sciences
Medicine and Health Sciences
Muscles
Myofibroblasts - cytology
Myofibroblasts - metabolism
Physical Sciences
Plates
Proteins
R&D
Research & development
Silicone resins
Silicones
Smooth muscle
Stiffness
Surgery
Transcription
Transcription Factors - metabolism
Transforming Growth Factor beta2 - metabolism
Transforming Growth Factor beta2 - pharmacology
Wound healing
Yes-associated protein
title TGF-β-induced activation of conjunctival fibroblasts is modulated by FGF-2 and substratum stiffness
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