AIM:To explore the effect of epidermal growth factor receptor(EGFR)inhibition by erlotinib and EGFR siRNA on epidermal growth factor(EGF)-induced activation of retinal pigment epithelium(RPE)cells.METHODS:Human RPE ce...AIM:To explore the effect of epidermal growth factor receptor(EGFR)inhibition by erlotinib and EGFR siRNA on epidermal growth factor(EGF)-induced activation of retinal pigment epithelium(RPE)cells.METHODS:Human RPE cell line(ARPE-19 cells)was activated by 100 ng/mL EGF.Erlotinib and EGFR siRNA were used to intervene EGF treatment.Cellular viability,proliferation,and migration were detected by methyl thiazolyl tetrazolium(MTT)assay,bromodeoxyuridine(BrdU)staining assay and wound healing assay,respectively.EGFR/protein kinase B(AKT)pathway proteins and N-cadherin,α-smooth muscle actin(α-SMA),and vimentin were tested by Western blot assay.EGFR was also determined by immunofluorescence staining.RESULTS:EGF treatment for 24h induced a significant increase of ARPE-19 cells’viability,proliferation and migration,phosphorylation of EGFR/AKT proteins,and decreased total EGFR expression.Erlotinib suppressed ARPE-19 cells’viability,proliferation and migration through down regulating total EGFR and AKT protein expressions.Erlotinib also inhibited EGF-induced an increase of proliferative and migrative ability in ARPE-19 cells and clearly suppressed EGF-induced EGFR/AKT proteins phosphorylation and decreased expression of N-cadherin,α-SMA,and vimentin proteins.Similarly,EGFR inhibition by EGFR siRNA significantly affected EGF-induced an increase of cell proliferation,viability,and migration,phosphorylation of EGFR/AKT proteins,and up-regulation of N-cadherin,α-SMA,and vimentin proteins.CONCLUSION:Erlotinib and EGFR-knockdown suppress EGF-induced cell viability,proliferation,and migration via EGFR/AKT pathway in RPE cells.EGFR inhibition may be a possible therapeutic approach for proliferative vitreoretinopathy(PVR).展开更多
Fibroblast growth factor receptors(FGFRs)play an important role in the regulation of cell proliferation,migration and differentiation,while the juxtamembrane domain(JMD)of FGFRs is the key in mediating these transmemb...Fibroblast growth factor receptors(FGFRs)play an important role in the regulation of cell proliferation,migration and differentiation,while the juxtamembrane domain(JMD)of FGFRs is the key in mediating these transmembrane signal transduction processes.Here,we expressed and purified the JMD(398K-470R)of FGFR1 with the presence of transmembrane domain(377I-397Y).The results from nuclear magnetic resonance(NMR)chemical shift analysis demonstrate that the main structure of JMD is disordered.Yet,the N-terminus of JMD was observed to form a short a-helix upon introducing negatively charged lipid 1,2-dioleoyl-sn-glycero-3-phospho-L-serine(DOPS)into its membrane mimic bicelles.Moreover,the N-terminus of JMD interacts with FRS2a,which is a substrate 2a of FGFR.Hence,we propose a model that FGFR1-JMD may interact with FRS2a and negatively charged lipids competitively.Our study provides a new understanding on the role of the JMD of FGFRs.展开更多
基金Supported by the Natural Science Foundation of Shaanxi Province,China(No.2022JM-521).
文摘AIM:To explore the effect of epidermal growth factor receptor(EGFR)inhibition by erlotinib and EGFR siRNA on epidermal growth factor(EGF)-induced activation of retinal pigment epithelium(RPE)cells.METHODS:Human RPE cell line(ARPE-19 cells)was activated by 100 ng/mL EGF.Erlotinib and EGFR siRNA were used to intervene EGF treatment.Cellular viability,proliferation,and migration were detected by methyl thiazolyl tetrazolium(MTT)assay,bromodeoxyuridine(BrdU)staining assay and wound healing assay,respectively.EGFR/protein kinase B(AKT)pathway proteins and N-cadherin,α-smooth muscle actin(α-SMA),and vimentin were tested by Western blot assay.EGFR was also determined by immunofluorescence staining.RESULTS:EGF treatment for 24h induced a significant increase of ARPE-19 cells’viability,proliferation and migration,phosphorylation of EGFR/AKT proteins,and decreased total EGFR expression.Erlotinib suppressed ARPE-19 cells’viability,proliferation and migration through down regulating total EGFR and AKT protein expressions.Erlotinib also inhibited EGF-induced an increase of proliferative and migrative ability in ARPE-19 cells and clearly suppressed EGF-induced EGFR/AKT proteins phosphorylation and decreased expression of N-cadherin,α-SMA,and vimentin proteins.Similarly,EGFR inhibition by EGFR siRNA significantly affected EGF-induced an increase of cell proliferation,viability,and migration,phosphorylation of EGFR/AKT proteins,and up-regulation of N-cadherin,α-SMA,and vimentin proteins.CONCLUSION:Erlotinib and EGFR-knockdown suppress EGF-induced cell viability,proliferation,and migration via EGFR/AKT pathway in RPE cells.EGFR inhibition may be a possible therapeutic approach for proliferative vitreoretinopathy(PVR).
基金Special Foundation of President of the Chinese Academy of Sciences(Grant No.,YZJJ2021QN33,YZJJ2020QN27)Collaborative Innovation Program of Hefei Science Center,CAS(2020HSC-CIP008).
文摘Fibroblast growth factor receptors(FGFRs)play an important role in the regulation of cell proliferation,migration and differentiation,while the juxtamembrane domain(JMD)of FGFRs is the key in mediating these transmembrane signal transduction processes.Here,we expressed and purified the JMD(398K-470R)of FGFR1 with the presence of transmembrane domain(377I-397Y).The results from nuclear magnetic resonance(NMR)chemical shift analysis demonstrate that the main structure of JMD is disordered.Yet,the N-terminus of JMD was observed to form a short a-helix upon introducing negatively charged lipid 1,2-dioleoyl-sn-glycero-3-phospho-L-serine(DOPS)into its membrane mimic bicelles.Moreover,the N-terminus of JMD interacts with FRS2a,which is a substrate 2a of FGFR.Hence,we propose a model that FGFR1-JMD may interact with FRS2a and negatively charged lipids competitively.Our study provides a new understanding on the role of the JMD of FGFRs.
