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(Pro)renin receptor is crucial for Wnt/β-catenin-dependent genesis of pancreatic ductal adenocarcinoma.

Shibayama Y, Fujimori T, Nguyen G, Hirose T, Totsune K, Ichihara A, Kitada K, Nakano D, Kobori H, Kohno M, Masaki T, Suzuki Y, Yachida S, Nishiyama A - Sci Rep (2015)

Bottom Line: Plasma s(P)RR levels were significantly (P < 0.0001) higher in patients with PDAC than in healthy matched controls.Loss of (P)RR induced apoptosis of human PDAC cells.This is the first demonstration that (P)RR may be profoundly involved in ductal tumorigenesis in the pancreas.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan.

ABSTRACT
Although Wnt/β-catenin signaling is known to be aberrantly activated in PDAC, mutations of CTNNB1, APC or other pathway components are rare in this tumor type, suggesting alternative mechanisms for Wnt/β-catenin activation. Recent studies have implicated the (pro)renin receptor ((P)RR) is related to the Wnt/β-catenin signaling pathway. We therefore investigated the possible role of (P)RR in pancreatic carcinogenesis. Plasma s(P)RR levels were significantly (P < 0.0001) higher in patients with PDAC than in healthy matched controls. We also identified aberrant expression of (P)RR in premalignant PanIN and PDAC lesions and all the PDAC cell lines examined. Inhibiting (P)RR with an siRNA attenuated activation of Wnt/β-catenin signaling pathway and reduced the proliferative ability of PDAC cells in vitro and the growth of engrafted tumors in vivo. Loss of (P)RR induced apoptosis of human PDAC cells. This is the first demonstration that (P)RR may be profoundly involved in ductal tumorigenesis in the pancreas.

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Related in: MedlinePlus

(P) RR is essential for activating the Wnt/β-catenin signaling pathway in human PDAC cell lines.(a) Representative image of Wnt3a expression among three human PDAC cell lines. Consistent results were observed when three experiments were repeated. (b) Wnt3a (150 ng/mL) significantly increased LRP6 activity in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. Blotting with an anti-LRP6 antibody showed equal loading. (c) Wnt3a significantly increased active β-catenin expression in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. β-actin was used as a loading control. (d) Effect of (P)RR siRNA on LRP6 activity in Wnt3a-treated PK-1 cells (mean ± SEM, n = 3 for each). Blotting with an anti-LRP6 antibody showed equal loading. (P)RR protein expression indicates efficient gene transfection. (e) Effect of (P)RR siRNA on active β-catenin and Cyclin D1 expression in PK-1, BxPC-3 and PANC-1 cells (mean ± SEM, n = 3 for each, *P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells; **P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells stimulated with Wnt3a; #P < 0.05 vs. CyclinD1 expression in scrambled siRNA-transfected cells; §P < 0.05 vs. Cyclin D1 expression of scrambled siRNA-transfected cells stimulated with Wnt3a). β-actin was used as a loading control.
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f3: (P) RR is essential for activating the Wnt/β-catenin signaling pathway in human PDAC cell lines.(a) Representative image of Wnt3a expression among three human PDAC cell lines. Consistent results were observed when three experiments were repeated. (b) Wnt3a (150 ng/mL) significantly increased LRP6 activity in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. Blotting with an anti-LRP6 antibody showed equal loading. (c) Wnt3a significantly increased active β-catenin expression in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. β-actin was used as a loading control. (d) Effect of (P)RR siRNA on LRP6 activity in Wnt3a-treated PK-1 cells (mean ± SEM, n = 3 for each). Blotting with an anti-LRP6 antibody showed equal loading. (P)RR protein expression indicates efficient gene transfection. (e) Effect of (P)RR siRNA on active β-catenin and Cyclin D1 expression in PK-1, BxPC-3 and PANC-1 cells (mean ± SEM, n = 3 for each, *P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells; **P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells stimulated with Wnt3a; #P < 0.05 vs. CyclinD1 expression in scrambled siRNA-transfected cells; §P < 0.05 vs. Cyclin D1 expression of scrambled siRNA-transfected cells stimulated with Wnt3a). β-actin was used as a loading control.

Mentions: We aimed to investigate whether (P)RR is a key factor for the canonical (β-catenin-dependent) Wnt signaling pathway in human PDAC cells. We examined the effect of Wnt3a, which is essentially involved in cell survival2930, as previously reported in studies of the pancreas1331. Endogenous Wnt3a expression was confirmed among three different PDAC cell lines. Wnt3a at basal conditions was highly expressed in PANC-1 cells (Fig. 3a). We focused on activation of Wnt3a-stimulated LRP6, which is a part of the Wnt receptor complex that also comprises (P)RR25, and on the non-phosphorylated form of β-catenin (“active” β-catenin) expression, which is thought to be responsible for mediating Wnt signaling in human PDAC cells32. In PK-1 cells, treatment with recombinant human Wnt3a (150 ng/mL) significantly enhanced phosphorylation of LRP6, a peak being reached after 10 min (Fig. 3b). Similarly, Wnt3a enhanced the expression of active β-catenin on and off from 10 min (Fig. 3c), suggesting that Wnt3a simultaneously induces phosphorylation of LRP6 and active β-catenin expression.


(Pro)renin receptor is crucial for Wnt/β-catenin-dependent genesis of pancreatic ductal adenocarcinoma.

Shibayama Y, Fujimori T, Nguyen G, Hirose T, Totsune K, Ichihara A, Kitada K, Nakano D, Kobori H, Kohno M, Masaki T, Suzuki Y, Yachida S, Nishiyama A - Sci Rep (2015)

(P) RR is essential for activating the Wnt/β-catenin signaling pathway in human PDAC cell lines.(a) Representative image of Wnt3a expression among three human PDAC cell lines. Consistent results were observed when three experiments were repeated. (b) Wnt3a (150 ng/mL) significantly increased LRP6 activity in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. Blotting with an anti-LRP6 antibody showed equal loading. (c) Wnt3a significantly increased active β-catenin expression in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. β-actin was used as a loading control. (d) Effect of (P)RR siRNA on LRP6 activity in Wnt3a-treated PK-1 cells (mean ± SEM, n = 3 for each). Blotting with an anti-LRP6 antibody showed equal loading. (P)RR protein expression indicates efficient gene transfection. (e) Effect of (P)RR siRNA on active β-catenin and Cyclin D1 expression in PK-1, BxPC-3 and PANC-1 cells (mean ± SEM, n = 3 for each, *P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells; **P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells stimulated with Wnt3a; #P < 0.05 vs. CyclinD1 expression in scrambled siRNA-transfected cells; §P < 0.05 vs. Cyclin D1 expression of scrambled siRNA-transfected cells stimulated with Wnt3a). β-actin was used as a loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4352858&req=5

f3: (P) RR is essential for activating the Wnt/β-catenin signaling pathway in human PDAC cell lines.(a) Representative image of Wnt3a expression among three human PDAC cell lines. Consistent results were observed when three experiments were repeated. (b) Wnt3a (150 ng/mL) significantly increased LRP6 activity in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. Blotting with an anti-LRP6 antibody showed equal loading. (c) Wnt3a significantly increased active β-catenin expression in PK-1 cells (mean ± SEM, n = 3 for each). *P < 0.05 vs. vehicle-treated cells. β-actin was used as a loading control. (d) Effect of (P)RR siRNA on LRP6 activity in Wnt3a-treated PK-1 cells (mean ± SEM, n = 3 for each). Blotting with an anti-LRP6 antibody showed equal loading. (P)RR protein expression indicates efficient gene transfection. (e) Effect of (P)RR siRNA on active β-catenin and Cyclin D1 expression in PK-1, BxPC-3 and PANC-1 cells (mean ± SEM, n = 3 for each, *P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells; **P < 0.05 vs. active β-catenin expression in scrambled siRNA-transfected cells stimulated with Wnt3a; #P < 0.05 vs. CyclinD1 expression in scrambled siRNA-transfected cells; §P < 0.05 vs. Cyclin D1 expression of scrambled siRNA-transfected cells stimulated with Wnt3a). β-actin was used as a loading control.
Mentions: We aimed to investigate whether (P)RR is a key factor for the canonical (β-catenin-dependent) Wnt signaling pathway in human PDAC cells. We examined the effect of Wnt3a, which is essentially involved in cell survival2930, as previously reported in studies of the pancreas1331. Endogenous Wnt3a expression was confirmed among three different PDAC cell lines. Wnt3a at basal conditions was highly expressed in PANC-1 cells (Fig. 3a). We focused on activation of Wnt3a-stimulated LRP6, which is a part of the Wnt receptor complex that also comprises (P)RR25, and on the non-phosphorylated form of β-catenin (“active” β-catenin) expression, which is thought to be responsible for mediating Wnt signaling in human PDAC cells32. In PK-1 cells, treatment with recombinant human Wnt3a (150 ng/mL) significantly enhanced phosphorylation of LRP6, a peak being reached after 10 min (Fig. 3b). Similarly, Wnt3a enhanced the expression of active β-catenin on and off from 10 min (Fig. 3c), suggesting that Wnt3a simultaneously induces phosphorylation of LRP6 and active β-catenin expression.

Bottom Line: Plasma s(P)RR levels were significantly (P < 0.0001) higher in patients with PDAC than in healthy matched controls.Loss of (P)RR induced apoptosis of human PDAC cells.This is the first demonstration that (P)RR may be profoundly involved in ductal tumorigenesis in the pancreas.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine, Kagawa University, Kagawa 761-0793, Japan.

ABSTRACT
Although Wnt/β-catenin signaling is known to be aberrantly activated in PDAC, mutations of CTNNB1, APC or other pathway components are rare in this tumor type, suggesting alternative mechanisms for Wnt/β-catenin activation. Recent studies have implicated the (pro)renin receptor ((P)RR) is related to the Wnt/β-catenin signaling pathway. We therefore investigated the possible role of (P)RR in pancreatic carcinogenesis. Plasma s(P)RR levels were significantly (P < 0.0001) higher in patients with PDAC than in healthy matched controls. We also identified aberrant expression of (P)RR in premalignant PanIN and PDAC lesions and all the PDAC cell lines examined. Inhibiting (P)RR with an siRNA attenuated activation of Wnt/β-catenin signaling pathway and reduced the proliferative ability of PDAC cells in vitro and the growth of engrafted tumors in vivo. Loss of (P)RR induced apoptosis of human PDAC cells. This is the first demonstration that (P)RR may be profoundly involved in ductal tumorigenesis in the pancreas.

Show MeSH
Related in: MedlinePlus