Rapamycin inhibits transforming growth factor beta 1 induced myofibroblast differentiation via the phosphorylated'phosphatidylinositol 3-kinase mammalian target of rapamycin signal pathways in nasal polyp'derived fibroblasts

Dong Yn Ko, Jae Min Shin, Ji Young Um, Byungjin Kang, Il Ho Park, Heung Man Lee

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Purpose: Rapamycin has antiproliferative and antifibrogenic effects in vitro and in vivo. The purpose of this study was to evaluate the effects of rapamycin on transforming growth factor (TGF) beta 1 induced myofibroblast differentiation (alpha smooth-muscle actin [SMA]), extracellular matrix production, and collagen contraction in nasal polyp' derived fibroblasts (NPDF). The underlying molecular mechanisms of rapamycin were also determined in NPDFs. Methods: NPDFs were grown in culture and transformed into myofibroblasts by using TGF beta 1 (5 ng/mL). For cytotoxicity evaluation, a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay was used. Expression levels of alpha SMA, phosphorylated phosphatidylinositol 3-kinase (PI3K), and phosphorylated mammalian target of rapamycin (mTOR) were determined by using Western blot, reverse transcription'polymerase chain reaction, and immunofluorescence staining. The total amount of collagen was analyzed by using the Sircol collagen assay, and contractile activity was measured with a collagen gel contraction assay. Silencing mTOR with mTOR-specific small interference RNA was determined by using reverse transcription'polymerase chain reaction. Results: Whereas rapamycin (range, 0'400 nM) had no significant cytotoxic effects on TGF beta 1 induced NPDFs, it significantly reduced the expression levels of alpha'SMA in TGF beta 1 induced NPDFs in a dose-dependent manner. TGF beta 1 induced collagen production and collagen contraction were significantly inhibited by rapamycin treatment. Rapamycin also attenuated the TGF beta 1 induced activation of PI3K and mTOR, and its inhibitory effects were similar to those of mTOR silencing and a specific PI3K inhibitor. Conclusions: Rapamycin inhibited TGF beta 1 induced myofibroblast differentiation, extracellular matrix production, and collagen contraction through the PI3K/mTOR signal pathways in NPDFs.

Original languageEnglish
Pages (from-to)e211-e217
JournalAmerican Journal of Rhinology and Allergy
Volume30
Issue number6
DOIs
Publication statusPublished - 2016 Nov 1

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Phosphatidylinositol 3-Kinase
Nasal Polyps
Myofibroblasts
Sirolimus
Transforming Growth Factor beta
Signal Transduction
Fibroblasts
Collagen
Smooth Muscle
Actins
Reverse Transcription
Extracellular Matrix
Polymerase Chain Reaction
RNA Interference
Bromides

ASJC Scopus subject areas

  • Immunology and Allergy
  • Otorhinolaryngology

Cite this

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title = "Rapamycin inhibits transforming growth factor beta 1 induced myofibroblast differentiation via the phosphorylated'phosphatidylinositol 3-kinase mammalian target of rapamycin signal pathways in nasal polyp'derived fibroblasts",
abstract = "Purpose: Rapamycin has antiproliferative and antifibrogenic effects in vitro and in vivo. The purpose of this study was to evaluate the effects of rapamycin on transforming growth factor (TGF) beta 1 induced myofibroblast differentiation (alpha smooth-muscle actin [SMA]), extracellular matrix production, and collagen contraction in nasal polyp' derived fibroblasts (NPDF). The underlying molecular mechanisms of rapamycin were also determined in NPDFs. Methods: NPDFs were grown in culture and transformed into myofibroblasts by using TGF beta 1 (5 ng/mL). For cytotoxicity evaluation, a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay was used. Expression levels of alpha SMA, phosphorylated phosphatidylinositol 3-kinase (PI3K), and phosphorylated mammalian target of rapamycin (mTOR) were determined by using Western blot, reverse transcription'polymerase chain reaction, and immunofluorescence staining. The total amount of collagen was analyzed by using the Sircol collagen assay, and contractile activity was measured with a collagen gel contraction assay. Silencing mTOR with mTOR-specific small interference RNA was determined by using reverse transcription'polymerase chain reaction. Results: Whereas rapamycin (range, 0'400 nM) had no significant cytotoxic effects on TGF beta 1 induced NPDFs, it significantly reduced the expression levels of alpha'SMA in TGF beta 1 induced NPDFs in a dose-dependent manner. TGF beta 1 induced collagen production and collagen contraction were significantly inhibited by rapamycin treatment. Rapamycin also attenuated the TGF beta 1 induced activation of PI3K and mTOR, and its inhibitory effects were similar to those of mTOR silencing and a specific PI3K inhibitor. Conclusions: Rapamycin inhibited TGF beta 1 induced myofibroblast differentiation, extracellular matrix production, and collagen contraction through the PI3K/mTOR signal pathways in NPDFs.",
author = "Ko, {Dong Yn} and Shin, {Jae Min} and Um, {Ji Young} and Byungjin Kang and Park, {Il Ho} and Lee, {Heung Man}",
year = "2016",
month = "11",
day = "1",
doi = "10.2500/ajra.2016.30.4389",
language = "English",
volume = "30",
pages = "e211--e217",
journal = "American Journal of Rhinology and Allergy",
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TY - JOUR

T1 - Rapamycin inhibits transforming growth factor beta 1 induced myofibroblast differentiation via the phosphorylated'phosphatidylinositol 3-kinase mammalian target of rapamycin signal pathways in nasal polyp'derived fibroblasts

AU - Ko, Dong Yn

AU - Shin, Jae Min

AU - Um, Ji Young

AU - Kang, Byungjin

AU - Park, Il Ho

AU - Lee, Heung Man

PY - 2016/11/1

Y1 - 2016/11/1

N2 - Purpose: Rapamycin has antiproliferative and antifibrogenic effects in vitro and in vivo. The purpose of this study was to evaluate the effects of rapamycin on transforming growth factor (TGF) beta 1 induced myofibroblast differentiation (alpha smooth-muscle actin [SMA]), extracellular matrix production, and collagen contraction in nasal polyp' derived fibroblasts (NPDF). The underlying molecular mechanisms of rapamycin were also determined in NPDFs. Methods: NPDFs were grown in culture and transformed into myofibroblasts by using TGF beta 1 (5 ng/mL). For cytotoxicity evaluation, a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay was used. Expression levels of alpha SMA, phosphorylated phosphatidylinositol 3-kinase (PI3K), and phosphorylated mammalian target of rapamycin (mTOR) were determined by using Western blot, reverse transcription'polymerase chain reaction, and immunofluorescence staining. The total amount of collagen was analyzed by using the Sircol collagen assay, and contractile activity was measured with a collagen gel contraction assay. Silencing mTOR with mTOR-specific small interference RNA was determined by using reverse transcription'polymerase chain reaction. Results: Whereas rapamycin (range, 0'400 nM) had no significant cytotoxic effects on TGF beta 1 induced NPDFs, it significantly reduced the expression levels of alpha'SMA in TGF beta 1 induced NPDFs in a dose-dependent manner. TGF beta 1 induced collagen production and collagen contraction were significantly inhibited by rapamycin treatment. Rapamycin also attenuated the TGF beta 1 induced activation of PI3K and mTOR, and its inhibitory effects were similar to those of mTOR silencing and a specific PI3K inhibitor. Conclusions: Rapamycin inhibited TGF beta 1 induced myofibroblast differentiation, extracellular matrix production, and collagen contraction through the PI3K/mTOR signal pathways in NPDFs.

AB - Purpose: Rapamycin has antiproliferative and antifibrogenic effects in vitro and in vivo. The purpose of this study was to evaluate the effects of rapamycin on transforming growth factor (TGF) beta 1 induced myofibroblast differentiation (alpha smooth-muscle actin [SMA]), extracellular matrix production, and collagen contraction in nasal polyp' derived fibroblasts (NPDF). The underlying molecular mechanisms of rapamycin were also determined in NPDFs. Methods: NPDFs were grown in culture and transformed into myofibroblasts by using TGF beta 1 (5 ng/mL). For cytotoxicity evaluation, a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay was used. Expression levels of alpha SMA, phosphorylated phosphatidylinositol 3-kinase (PI3K), and phosphorylated mammalian target of rapamycin (mTOR) were determined by using Western blot, reverse transcription'polymerase chain reaction, and immunofluorescence staining. The total amount of collagen was analyzed by using the Sircol collagen assay, and contractile activity was measured with a collagen gel contraction assay. Silencing mTOR with mTOR-specific small interference RNA was determined by using reverse transcription'polymerase chain reaction. Results: Whereas rapamycin (range, 0'400 nM) had no significant cytotoxic effects on TGF beta 1 induced NPDFs, it significantly reduced the expression levels of alpha'SMA in TGF beta 1 induced NPDFs in a dose-dependent manner. TGF beta 1 induced collagen production and collagen contraction were significantly inhibited by rapamycin treatment. Rapamycin also attenuated the TGF beta 1 induced activation of PI3K and mTOR, and its inhibitory effects were similar to those of mTOR silencing and a specific PI3K inhibitor. Conclusions: Rapamycin inhibited TGF beta 1 induced myofibroblast differentiation, extracellular matrix production, and collagen contraction through the PI3K/mTOR signal pathways in NPDFs.

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U2 - 10.2500/ajra.2016.30.4389

DO - 10.2500/ajra.2016.30.4389

M3 - Article

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JO - American Journal of Rhinology and Allergy

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SN - 1945-8924

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