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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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Wild-type β-catenin or constitutively active mutant β-catenin constructs were cotransfected into PK-1 cells with scrambled siRNA or (P)RR siRNA expression.(a) Effect of wild-type β-catenin (wt β-catenin) on the (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. (b) The protein expression of (P)RR and Myc indicates efficiency of gene transfection with (P)RR (ATP6AP2) siRNA and a plasmid carrying a constitutively active aa 1–90-deleted β-catenin tagged with Myc, respectively. (c) Effect of constitutively active mutant β-catenin (mut β-catenin) on (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. Cell proliferation was evaluated at 24 h and 48 h after Wnt 3a stimulation by WST-1 assays (mean ± SEM, n = 3 for each, P < 0.05 vs. (P)RR siRNA cells). N.S., not significant.
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f5: Wild-type β-catenin or constitutively active mutant β-catenin constructs were cotransfected into PK-1 cells with scrambled siRNA or (P)RR siRNA expression.(a) Effect of wild-type β-catenin (wt β-catenin) on the (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. (b) The protein expression of (P)RR and Myc indicates efficiency of gene transfection with (P)RR (ATP6AP2) siRNA and a plasmid carrying a constitutively active aa 1–90-deleted β-catenin tagged with Myc, respectively. (c) Effect of constitutively active mutant β-catenin (mut β-catenin) on (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. Cell proliferation was evaluated at 24 h and 48 h after Wnt 3a stimulation by WST-1 assays (mean ± SEM, n = 3 for each, P < 0.05 vs. (P)RR siRNA cells). N.S., not significant.

Mentions: To investigate whether inhibition of (P)RR reduces Wnt3a-induced PDAC cell proliferation through inactivation of β-catenin, we examined the effects of co-transfection of (P)RR siRNA and a plasmid harboring wild-type β-catenin or constitutive activation of mutant β-catenin on Wnt3a-induced cell proliferative ability. Consistent with the above-mentioned data (Fig. 4a), treatment with siRNA against (P)RR significantly decreased the proliferative ability of Wnt3a-stimulated PK-1 cells (Fig. 5a–c). In these cells, overexpression of wild-type β-catenin did not affect the siRNA (P)RR-induced reduction in cell proliferative ability (Fig. 5a). Compared with cells transfected with (P)RR siRNA, constitutive activation of β-catenin significantly maintained the proliferative ability of (P)RR siRNA-transfected cells after 24 h by Wnt3a stimulation (Fig. 5b, c) but resulted in the same effect as (P)RR siRNA knockdown after 48 h (Fig. 5c). Thus, these results indicate that transfection by constitutive activation of β-catenin contributes to a delay in decreasing the proliferative ability of (P)RR siRNA-transfected cells, but the siRNA (P)RR-induced reduction in cell proliferative ability is not fully recovered through transfection with constitutive activation of β-catenin. These data confirm that (P)RR function is upstream of β-catenin in human PDAC cells, as recently demonstrated in Xenopus embryos25, but the decreased proliferative ability of (P)RR siRNA-transfected cells cannot be explained simply by inactivation of the Wnt/β-catenin signaling cascade.


(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)

Wild-type β-catenin or constitutively active mutant β-catenin constructs were cotransfected into PK-1 cells with scrambled siRNA or (P)RR siRNA expression.(a) Effect of wild-type β-catenin (wt β-catenin) on the (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. (b) The protein expression of (P)RR and Myc indicates efficiency of gene transfection with (P)RR (ATP6AP2) siRNA and a plasmid carrying a constitutively active aa 1–90-deleted β-catenin tagged with Myc, respectively. (c) Effect of constitutively active mutant β-catenin (mut β-catenin) on (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. Cell proliferation was evaluated at 24 h and 48 h after Wnt 3a stimulation by WST-1 assays (mean ± SEM, n = 3 for each, P < 0.05 vs. (P)RR siRNA cells). N.S., not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4352858&req=5

f5: Wild-type β-catenin or constitutively active mutant β-catenin constructs were cotransfected into PK-1 cells with scrambled siRNA or (P)RR siRNA expression.(a) Effect of wild-type β-catenin (wt β-catenin) on the (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. (b) The protein expression of (P)RR and Myc indicates efficiency of gene transfection with (P)RR (ATP6AP2) siRNA and a plasmid carrying a constitutively active aa 1–90-deleted β-catenin tagged with Myc, respectively. (c) Effect of constitutively active mutant β-catenin (mut β-catenin) on (P)RR siRNA-induced reduction in PK-1 cell proliferative ability. Cell proliferation was evaluated at 24 h and 48 h after Wnt 3a stimulation by WST-1 assays (mean ± SEM, n = 3 for each, P < 0.05 vs. (P)RR siRNA cells). N.S., not significant.
Mentions: To investigate whether inhibition of (P)RR reduces Wnt3a-induced PDAC cell proliferation through inactivation of β-catenin, we examined the effects of co-transfection of (P)RR siRNA and a plasmid harboring wild-type β-catenin or constitutive activation of mutant β-catenin on Wnt3a-induced cell proliferative ability. Consistent with the above-mentioned data (Fig. 4a), treatment with siRNA against (P)RR significantly decreased the proliferative ability of Wnt3a-stimulated PK-1 cells (Fig. 5a–c). In these cells, overexpression of wild-type β-catenin did not affect the siRNA (P)RR-induced reduction in cell proliferative ability (Fig. 5a). Compared with cells transfected with (P)RR siRNA, constitutive activation of β-catenin significantly maintained the proliferative ability of (P)RR siRNA-transfected cells after 24 h by Wnt3a stimulation (Fig. 5b, c) but resulted in the same effect as (P)RR siRNA knockdown after 48 h (Fig. 5c). Thus, these results indicate that transfection by constitutive activation of β-catenin contributes to a delay in decreasing the proliferative ability of (P)RR siRNA-transfected cells, but the siRNA (P)RR-induced reduction in cell proliferative ability is not fully recovered through transfection with constitutive activation of β-catenin. These data confirm that (P)RR function is upstream of β-catenin in human PDAC cells, as recently demonstrated in Xenopus embryos25, but the decreased proliferative ability of (P)RR siRNA-transfected cells cannot be explained simply by inactivation of the Wnt/β-catenin signaling cascade.

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