TY - JOUR
T1 - GPR75 receptor mediates 20-HETE-signaling and metastatic features of androgen-insensitive prostate cancer cells
AU - Cárdenas, Sofia
AU - Colombero, Cecilia
AU - Panelo, Laura
AU - Dakarapu, Rambabu
AU - Falck, John R.
AU - Costas, Monica A.
AU - Nowicki, Susana
N1 - Funding Information:
This work was supported by grant 1006/2016 from the National Cancer Institute (INC-Argentina) to SN, grant 11220150100118CO from the Argentinean National Research Council (CONICET) to MC and SN. JRF was supported by the Robert A. Welch Foundation (I-0011) and NIH HL139793.
Funding Information:
This work was supported by grant 1006/2016 from the National Cancer Institute (INC-Argentina) to SN, grant 11220150100118CO from the Argentinean National Research Council ( CONICET ) to MC and SN. JRF was supported by the Robert A. Welch Foundation ( I-0011 ) and NIH HL139793 .
Publisher Copyright:
© 2019 Elsevier B.V.
PY - 2020/2
Y1 - 2020/2
N2 - Purpose: Recent studies have shown that 20-hydroxyeicosatetraenoic acid (20-HETE) is a key molecule in sustaining androgen-mediated prostate cancer cell survival. Thus, the aim of this study was to determine whether 20-HETE can affect the metastatic potential of androgen-insensitive prostate cancer cells, and the implication of the newly described 20-HETE receptor, GPR75, in mediating these effects. Methods: The expression of GPR75, protein phosphorylation, actin polymerization and protein distribution were assessed by western blot and/or fluorescence microscopy. Additionally, in vitro assays including epithelial-mesenchymal transition (EMT), metalloproteinase-2 (MMP-2) activity, scratch wound healing, transwell invasion and soft agar colony formation were used to evaluate the effects of 20-HETE agonists/antagonists or GPR75 gene silencing on the aggressive features of PC-3 cells. Results: 20-HETE (0.1 nM) promoted the acquisition of a mesenchymal phenotype by increasing EMT, the release of MMP-2, cell migration and invasion, actin stress fiber formation and anchorage-independent growth. Also, 20-HETE augmented the expression of HIC-5, the phosphorylation of EGFR, NF-κB, AKT and p-38 and the intracellular redistribution of p-AKT and PKCα. These effects were impaired by GPR75 antagonism and/or silencing. Accordingly, the inhibition of 20-HETE formation with N-hydroxy-N′-(4-n-butyl-2-methylphenyl) formamidine (HET0016) elicited the opposite effects. Conclusions: The present results show for the first time the involvement of the 20-HETE-GPR75 receptor in the activation of intracellular signaling known to be stimulated in cell malignant transformations leading to the differentiation of PC-3 cells towards a more aggressive phenotype. Targeting the 20-HETE/GPR75 pathway is a promising and novel approach to interfere with prostate tumor cell malignant progression.
AB - Purpose: Recent studies have shown that 20-hydroxyeicosatetraenoic acid (20-HETE) is a key molecule in sustaining androgen-mediated prostate cancer cell survival. Thus, the aim of this study was to determine whether 20-HETE can affect the metastatic potential of androgen-insensitive prostate cancer cells, and the implication of the newly described 20-HETE receptor, GPR75, in mediating these effects. Methods: The expression of GPR75, protein phosphorylation, actin polymerization and protein distribution were assessed by western blot and/or fluorescence microscopy. Additionally, in vitro assays including epithelial-mesenchymal transition (EMT), metalloproteinase-2 (MMP-2) activity, scratch wound healing, transwell invasion and soft agar colony formation were used to evaluate the effects of 20-HETE agonists/antagonists or GPR75 gene silencing on the aggressive features of PC-3 cells. Results: 20-HETE (0.1 nM) promoted the acquisition of a mesenchymal phenotype by increasing EMT, the release of MMP-2, cell migration and invasion, actin stress fiber formation and anchorage-independent growth. Also, 20-HETE augmented the expression of HIC-5, the phosphorylation of EGFR, NF-κB, AKT and p-38 and the intracellular redistribution of p-AKT and PKCα. These effects were impaired by GPR75 antagonism and/or silencing. Accordingly, the inhibition of 20-HETE formation with N-hydroxy-N′-(4-n-butyl-2-methylphenyl) formamidine (HET0016) elicited the opposite effects. Conclusions: The present results show for the first time the involvement of the 20-HETE-GPR75 receptor in the activation of intracellular signaling known to be stimulated in cell malignant transformations leading to the differentiation of PC-3 cells towards a more aggressive phenotype. Targeting the 20-HETE/GPR75 pathway is a promising and novel approach to interfere with prostate tumor cell malignant progression.
KW - 20-HETE
KW - Arachidonic acid metabolites
KW - GPR75 receptor
KW - Metastasis
KW - Prostate cancer
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U2 - 10.1016/j.bbalip.2019.158573
DO - 10.1016/j.bbalip.2019.158573
M3 - Article
C2 - 31760076
AN - SCOPUS:85076021934
SN - 1388-1981
VL - 1865
JO - BBA - Specialised Section On Lipids and Related Subjects
JF - BBA - Specialised Section On Lipids and Related Subjects
IS - 2
M1 - 158573
ER -