TY - JOUR
T1 - Estrogen receptor beta signaling inhibits PDGF induced human airway smooth muscle proliferation
AU - Ambhore, Nilesh Sudhakar
AU - Katragadda, Rathnavali
AU - Raju Kalidhindi, Rama Satyanarayana
AU - Thompson, Michael A.
AU - Pabelick, Christina M.
AU - Prakash, Y. S.
AU - Sathish, Venkatachalem
N1 - Funding Information:
Supported by NIH grants R01 HL123494 , R01 HL123494-02S1 (Venkatachalem), and R01 HL 088029 (Prakash). Additional support in part from ND EPSCoR with NSF #1355466 and NDSU RCA Activity . Authors would like to thank Dr. Jagadish Loganathan, Department of Pharmaceutical Sciences, NDSU for providing technical support in completion of this study. The authors also acknowledge Dr. Tao Wang, Core Biology Facility for helping in performing the flow cytometry study. Funding for the Core Biology Facility used in the publication was made possible by NIH Grant P30 GM103332-01 .
Funding Information:
Supported by NIH grants R01 HL123494, R01 HL123494-02S1 (Venkatachalem), and R01 HL 088029 (Prakash). Additional support in part from ND EPSCoR with NSF #1355466 and NDSU RCA Activity. Authors would like to thank Dr. Jagadish Loganathan, Department of Pharmaceutical Sciences, NDSU for providing technical support in completion of this study. The authors also acknowledge Dr. Tao Wang, Core Biology Facility for helping in performing the flow cytometry study. Funding for the Core Biology Facility used in the publication was made possible by NIH Grant P30 GM103332-01.
Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/11/15
Y1 - 2018/11/15
N2 - Airway smooth muscle (ASM) cell hyperplasia driven by persistent inflammation is a hallmark feature of remodeling in asthma. Sex steroid signaling in the lungs is of considerable interest, given epidemiological data showing more asthma in pre-menopausal women and aging men. Our previous studies demonstrated that estrogen receptor (ER) expression increases in asthmatic human ASM; however, very limited data are available regarding differential roles of ERα vs. ERβ isoforms in human ASM cell proliferation. In this study, we evaluated the effect of selective ERα and ERβ modulators on platelet-derived growth factor (PDGF)-stimulated ASM proliferation and the mechanisms involved. Asthmatic and non-asthmatic primary human ASM cells were treated with PDGF, 17β-estradiol, ERα-agonist and/or ERβ-agonist and/or G-protein-coupled estrogen receptor 30 (GPR30/GPER) agonist and proliferation was measured using MTT and CyQuant assays followed by cell cycle analysis. Transfection of small interfering RNA (siRNA) ERα and ERβ significantly altered the human ASM proliferation. The specificity of siRNA transfection was confirmed by Western blot analysis. Gene and protein expression of cell cycle-related antigens (PCNA and Ki67) and C/EBP were measured by RT-PCR and Western analysis, along with cell signaling proteins. PDGF significantly increased ASM proliferation in non-asthmatic and asthmatic cells. Treatment with PPT showed no significant effect on PDGF-induced proliferation, whereas WAY interestingly suppressed proliferation via inhibition of ERK1/2, Akt, and p38 signaling. PDGF-induced gene expression of PCNA, Ki67 and C/EBP in human ASM was significantly lower in cells pre-treated with WAY. Furthermore, WAY also inhibited PDGF-activated PCNA, C/EBP, cyclin-D1, and cyclin-E. Overall, we demonstrate ER isoform-specific signaling in the context of ASM proliferation. Activation of ERβ can diminish remodeling in human ASM by inhibiting pro-proliferative signaling pathways, and may point to a novel perception for blunting airway remodeling.
AB - Airway smooth muscle (ASM) cell hyperplasia driven by persistent inflammation is a hallmark feature of remodeling in asthma. Sex steroid signaling in the lungs is of considerable interest, given epidemiological data showing more asthma in pre-menopausal women and aging men. Our previous studies demonstrated that estrogen receptor (ER) expression increases in asthmatic human ASM; however, very limited data are available regarding differential roles of ERα vs. ERβ isoforms in human ASM cell proliferation. In this study, we evaluated the effect of selective ERα and ERβ modulators on platelet-derived growth factor (PDGF)-stimulated ASM proliferation and the mechanisms involved. Asthmatic and non-asthmatic primary human ASM cells were treated with PDGF, 17β-estradiol, ERα-agonist and/or ERβ-agonist and/or G-protein-coupled estrogen receptor 30 (GPR30/GPER) agonist and proliferation was measured using MTT and CyQuant assays followed by cell cycle analysis. Transfection of small interfering RNA (siRNA) ERα and ERβ significantly altered the human ASM proliferation. The specificity of siRNA transfection was confirmed by Western blot analysis. Gene and protein expression of cell cycle-related antigens (PCNA and Ki67) and C/EBP were measured by RT-PCR and Western analysis, along with cell signaling proteins. PDGF significantly increased ASM proliferation in non-asthmatic and asthmatic cells. Treatment with PPT showed no significant effect on PDGF-induced proliferation, whereas WAY interestingly suppressed proliferation via inhibition of ERK1/2, Akt, and p38 signaling. PDGF-induced gene expression of PCNA, Ki67 and C/EBP in human ASM was significantly lower in cells pre-treated with WAY. Furthermore, WAY also inhibited PDGF-activated PCNA, C/EBP, cyclin-D1, and cyclin-E. Overall, we demonstrate ER isoform-specific signaling in the context of ASM proliferation. Activation of ERβ can diminish remodeling in human ASM by inhibiting pro-proliferative signaling pathways, and may point to a novel perception for blunting airway remodeling.
KW - Asthma
KW - ERα receptor
KW - Estrogen
KW - Lung
KW - PCNA
KW - Sex steroids
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U2 - 10.1016/j.mce.2018.04.007
DO - 10.1016/j.mce.2018.04.007
M3 - Article
C2 - 29680290
AN - SCOPUS:85046134519
SN - 0303-7207
VL - 476
SP - 37
EP - 47
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
ER -