Limits...
GPER signalling in both cancer-associated fibroblasts and breast cancer cells mediates a feedforward IL1β/IL1R1 response.

De Marco P, Lappano R, De Francesco EM, Cirillo F, Pupo M, Avino S, Vivacqua A, Abonante S, Picard D, Maggiolini M - Sci Rep (2016)

Bottom Line: Cancer-associated fibroblasts (CAFs) contribute to the malignant aggressiveness through secreted factors like IL1β, which may drive pro-tumorigenic inflammatory phenotypes mainly acting via the cognate receptor named IL1R1.Thereby, ligand-activation of GPER generates a feedforward loop coupling IL1β induction by CAFs to IL1R1 expression by cancer cells, promoting the up-regulation of IL1β/IL1R1 target genes such as PTGES, COX2, RAGE and ABCG2.This regulatory interaction between the two cell types induces migration and invasive features in breast cancer cells including fibroblastoid cytoarchitecture and F-actin reorganization.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy and Health and Nutritional Sciences, University of Calabria, 87036 Rende, Italy.

ABSTRACT
Cancer-associated fibroblasts (CAFs) contribute to the malignant aggressiveness through secreted factors like IL1β, which may drive pro-tumorigenic inflammatory phenotypes mainly acting via the cognate receptor named IL1R1. Here, we demonstrate that signalling mediated by the G protein estrogen receptor (GPER) triggers IL1β and IL1R1 expression in CAFs and breast cancer cells, respectively. Thereby, ligand-activation of GPER generates a feedforward loop coupling IL1β induction by CAFs to IL1R1 expression by cancer cells, promoting the up-regulation of IL1β/IL1R1 target genes such as PTGES, COX2, RAGE and ABCG2. This regulatory interaction between the two cell types induces migration and invasive features in breast cancer cells including fibroblastoid cytoarchitecture and F-actin reorganization. A better understanding of the mechanisms involved in the regulation of pro-inflammatory cytokines by GPER-integrated estrogen signals may be useful to target these stroma-cancer interactions.

No MeSH data available.


Related in: MedlinePlus

GPER mediates the up-regulation of IL1R1 expression by E2 and G-1 in SkBr3 and MCF-7 breast cancer cells.(A) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting SkBr3 cells for 24 h with shGPER. (B) Efficacy of GPER silencing. (C) The induction of IL1R1 protein expression observed treating SkBr3 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. (D) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting MCF-7 cells for 24 h with shGPER. (E) Efficacy of GPER silencing. (F) The induction of IL1R1 protein expression observed treating MCF-7 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. IL1R1 protein levels in SkBr3 cells treated for 8 h with 10 nM E2 (G) and 100 nM G-1 (H) alone or in combination with 1 μM EGFR inhibitor AG1478 (AG), 1 μM MEK inhibitor PD98059 (PD), 1 μM PKC inhibitor GF109203X (GF), 1  μM PI3K inhibitor LY294,002 (LY), 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB 203580 (SB). IL1R1 protein levels in MCF-7 cells treated for 8 h with 10 nM E2 (I) and 100 nM G-1 (J) alone or in combination with 1 μM EGFR inhibitor AG, 1 μM MEK inhibitor PD, 1 μM PKC inhibitor GF, 1 μM PI3K inhibitor LY, 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB. β-actin serves as a loading control. Results shown are representative of at least two independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4829876&req=5

f3: GPER mediates the up-regulation of IL1R1 expression by E2 and G-1 in SkBr3 and MCF-7 breast cancer cells.(A) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting SkBr3 cells for 24 h with shGPER. (B) Efficacy of GPER silencing. (C) The induction of IL1R1 protein expression observed treating SkBr3 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. (D) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting MCF-7 cells for 24 h with shGPER. (E) Efficacy of GPER silencing. (F) The induction of IL1R1 protein expression observed treating MCF-7 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. IL1R1 protein levels in SkBr3 cells treated for 8 h with 10 nM E2 (G) and 100 nM G-1 (H) alone or in combination with 1 μM EGFR inhibitor AG1478 (AG), 1 μM MEK inhibitor PD98059 (PD), 1 μM PKC inhibitor GF109203X (GF), 1  μM PI3K inhibitor LY294,002 (LY), 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB 203580 (SB). IL1R1 protein levels in MCF-7 cells treated for 8 h with 10 nM E2 (I) and 100 nM G-1 (J) alone or in combination with 1 μM EGFR inhibitor AG, 1 μM MEK inhibitor PD, 1 μM PKC inhibitor GF, 1 μM PI3K inhibitor LY, 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB. β-actin serves as a loading control. Results shown are representative of at least two independent experiments.

Mentions: Pro-inflammatory factors secreted within the breast tumor microenvironment mainly act via cognate receptors expressed by cancer cells32. On the basis of the abovementioned results and previous studies showing that estrogens may regulate the levels of IL1R133, we evaluated whether GPER mediates IL1R1 expression in breast tumor cells. As shown in Fig. 2, E2 and G-1 up-regulated the mRNA (Fig. 2A,B) and protein expression (Fig. 2C–F) of IL1R1 in both SkBr3 and MCF-7 cells. Moreover, IL1R1 protein induction by E2 and G-1 was abolished knocking-down the expression of GPER as well as in the presence of the GPER antagonist G-15 in SkBr3 and MCF-7 cells (Fig. 3A–F). Next, the up-regulation of IL1R1 by E2 and G-1 was prevented using the EGFR inhibitor AG, the MEK inhibitor PD and the PKC inhibitor GF, while the inhibitors of PI3K, PKA and p38 transduction pathways namely LY, H89 and SB, respectively, did not show any effect (Fig. 3G–J) as observed using also the ER antagonist ICI (Supplementary Fig. 1). Altogether, these results suggest that E2 and G-1 trigger the up-regulation of IL1R1 in breast cancer cells through GPER-mediated signalling.


GPER signalling in both cancer-associated fibroblasts and breast cancer cells mediates a feedforward IL1β/IL1R1 response.

De Marco P, Lappano R, De Francesco EM, Cirillo F, Pupo M, Avino S, Vivacqua A, Abonante S, Picard D, Maggiolini M - Sci Rep (2016)

GPER mediates the up-regulation of IL1R1 expression by E2 and G-1 in SkBr3 and MCF-7 breast cancer cells.(A) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting SkBr3 cells for 24 h with shGPER. (B) Efficacy of GPER silencing. (C) The induction of IL1R1 protein expression observed treating SkBr3 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. (D) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting MCF-7 cells for 24 h with shGPER. (E) Efficacy of GPER silencing. (F) The induction of IL1R1 protein expression observed treating MCF-7 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. IL1R1 protein levels in SkBr3 cells treated for 8 h with 10 nM E2 (G) and 100 nM G-1 (H) alone or in combination with 1 μM EGFR inhibitor AG1478 (AG), 1 μM MEK inhibitor PD98059 (PD), 1 μM PKC inhibitor GF109203X (GF), 1  μM PI3K inhibitor LY294,002 (LY), 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB 203580 (SB). IL1R1 protein levels in MCF-7 cells treated for 8 h with 10 nM E2 (I) and 100 nM G-1 (J) alone or in combination with 1 μM EGFR inhibitor AG, 1 μM MEK inhibitor PD, 1 μM PKC inhibitor GF, 1 μM PI3K inhibitor LY, 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB. β-actin serves as a loading control. Results shown are representative of at least two independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829876&req=5

f3: GPER mediates the up-regulation of IL1R1 expression by E2 and G-1 in SkBr3 and MCF-7 breast cancer cells.(A) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting SkBr3 cells for 24 h with shGPER. (B) Efficacy of GPER silencing. (C) The induction of IL1R1 protein expression observed treating SkBr3 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. (D) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting MCF-7 cells for 24 h with shGPER. (E) Efficacy of GPER silencing. (F) The induction of IL1R1 protein expression observed treating MCF-7 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. IL1R1 protein levels in SkBr3 cells treated for 8 h with 10 nM E2 (G) and 100 nM G-1 (H) alone or in combination with 1 μM EGFR inhibitor AG1478 (AG), 1 μM MEK inhibitor PD98059 (PD), 1 μM PKC inhibitor GF109203X (GF), 1  μM PI3K inhibitor LY294,002 (LY), 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB 203580 (SB). IL1R1 protein levels in MCF-7 cells treated for 8 h with 10 nM E2 (I) and 100 nM G-1 (J) alone or in combination with 1 μM EGFR inhibitor AG, 1 μM MEK inhibitor PD, 1 μM PKC inhibitor GF, 1 μM PI3K inhibitor LY, 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB. β-actin serves as a loading control. Results shown are representative of at least two independent experiments.
Mentions: Pro-inflammatory factors secreted within the breast tumor microenvironment mainly act via cognate receptors expressed by cancer cells32. On the basis of the abovementioned results and previous studies showing that estrogens may regulate the levels of IL1R133, we evaluated whether GPER mediates IL1R1 expression in breast tumor cells. As shown in Fig. 2, E2 and G-1 up-regulated the mRNA (Fig. 2A,B) and protein expression (Fig. 2C–F) of IL1R1 in both SkBr3 and MCF-7 cells. Moreover, IL1R1 protein induction by E2 and G-1 was abolished knocking-down the expression of GPER as well as in the presence of the GPER antagonist G-15 in SkBr3 and MCF-7 cells (Fig. 3A–F). Next, the up-regulation of IL1R1 by E2 and G-1 was prevented using the EGFR inhibitor AG, the MEK inhibitor PD and the PKC inhibitor GF, while the inhibitors of PI3K, PKA and p38 transduction pathways namely LY, H89 and SB, respectively, did not show any effect (Fig. 3G–J) as observed using also the ER antagonist ICI (Supplementary Fig. 1). Altogether, these results suggest that E2 and G-1 trigger the up-regulation of IL1R1 in breast cancer cells through GPER-mediated signalling.

Bottom Line: Cancer-associated fibroblasts (CAFs) contribute to the malignant aggressiveness through secreted factors like IL1β, which may drive pro-tumorigenic inflammatory phenotypes mainly acting via the cognate receptor named IL1R1.Thereby, ligand-activation of GPER generates a feedforward loop coupling IL1β induction by CAFs to IL1R1 expression by cancer cells, promoting the up-regulation of IL1β/IL1R1 target genes such as PTGES, COX2, RAGE and ABCG2.This regulatory interaction between the two cell types induces migration and invasive features in breast cancer cells including fibroblastoid cytoarchitecture and F-actin reorganization.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy and Health and Nutritional Sciences, University of Calabria, 87036 Rende, Italy.

ABSTRACT
Cancer-associated fibroblasts (CAFs) contribute to the malignant aggressiveness through secreted factors like IL1β, which may drive pro-tumorigenic inflammatory phenotypes mainly acting via the cognate receptor named IL1R1. Here, we demonstrate that signalling mediated by the G protein estrogen receptor (GPER) triggers IL1β and IL1R1 expression in CAFs and breast cancer cells, respectively. Thereby, ligand-activation of GPER generates a feedforward loop coupling IL1β induction by CAFs to IL1R1 expression by cancer cells, promoting the up-regulation of IL1β/IL1R1 target genes such as PTGES, COX2, RAGE and ABCG2. This regulatory interaction between the two cell types induces migration and invasive features in breast cancer cells including fibroblastoid cytoarchitecture and F-actin reorganization. A better understanding of the mechanisms involved in the regulation of pro-inflammatory cytokines by GPER-integrated estrogen signals may be useful to target these stroma-cancer interactions.

No MeSH data available.


Related in: MedlinePlus