Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1
INTRODUCTION: Wnt/β-catenin signaling activation has been reported only during the late steps of Barrett’s esophagus (BE) neoplastic progression, but not in BE metaplasia, based on the absence of nuclear β-catenin. However, β-catenin transcriptional activity has been recorded in absence of robust nu...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-07-01
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| Series: | Neoplasia: An International Journal for Oncology Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1476558615000871 |
| _version_ | 1818366716999106560 |
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| author | Orestis Lyros Parvaneh Rafiee Linghui Nie Rituparna Medda Nebojsa Jovanovic Mary F. Otterson Behnaz Behmaram Ιnes Gockel Alexander Mackinnon Reza Shaker |
| author_facet | Orestis Lyros Parvaneh Rafiee Linghui Nie Rituparna Medda Nebojsa Jovanovic Mary F. Otterson Behnaz Behmaram Ιnes Gockel Alexander Mackinnon Reza Shaker |
| author_sort | Orestis Lyros |
| collection | DOAJ |
| description | INTRODUCTION: Wnt/β-catenin signaling activation has been reported only during the late steps of Barrett’s esophagus (BE) neoplastic progression, but not in BE metaplasia, based on the absence of nuclear β-catenin. However, β-catenin transcriptional activity has been recorded in absence of robust nuclear accumulation. Thus, we aimed to investigate the Wnt/β-catenin signaling in nondysplastic BE. METHODS: Esophageal tissues from healthy and BE patients without dysplasia were analyzed for Wnt target gene expression by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. Esophageal squamous (EPC1-& EPC2-hTERT), BE metaplastic (CP-A), and adenocarcinoma (OE33) cell lines were characterized for Wnt activation by qRT-PCR, Western blot, and luciferase assay. Wnt activity regulation was examined by using recombinant Wnt3a and Dickkopf-1 (Dkk1) as well as Dkk1 short interfering RNA. RESULTS: Wnt target genes (AXIN2, c-MYC, Cyclin D1, Dkk1) and Wnt3a were significantly upregulated in nondysplastic BE compared with squamous mucosa. Elevated levels of dephosphorylated β-catenin were detected in nondysplastic BE. Nuclear active β-catenin and TOPflash activity were increased in CP-A and OE33 cells compared with squamous cells. Wnt3a-mediated β-catenin signaling activation was abolished by Dkk1 in CP-A cells. TOPFlash activity was elevated following Dkk1 silencing in CP-A but not in OE33 cells. Dysplastic and esophageal adenocarcinoma tissues demonstrated further Dkk1 and AXIN2 overexpression. CONCLUSIONS: Despite the absence of robust nuclear accumulation, β-catenin is transcriptionally active in nondysplastic BE. Dkk1 overexpression regulates β-catenin signaling in BE metaplastic but not in adenocarcinoma cells, suggesting that early perturbation of Dkk1-mediated signaling suppression may contribute to BE malignant transformation. |
| first_indexed | 2024-12-13T22:40:35Z |
| format | Article |
| id | doaj.art-0cf616cc98544b5aaedb54a0ab130474 |
| institution | Directory Open Access Journal |
| issn | 1476-5586 1522-8002 |
| language | English |
| last_indexed | 2024-12-13T22:40:35Z |
| publishDate | 2015-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neoplasia: An International Journal for Oncology Research |
| spelling | doaj.art-0cf616cc98544b5aaedb54a0ab1304742022-12-21T23:28:51ZengElsevierNeoplasia: An International Journal for Oncology Research1476-55861522-80022015-07-0117759861110.1016/j.neo.2015.07.006Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1Orestis Lyros0Parvaneh Rafiee1Linghui Nie2Rituparna Medda3Nebojsa Jovanovic4Mary F. Otterson5Behnaz Behmaram6Ιnes Gockel7Alexander Mackinnon8Reza Shaker9Division of Gastroenterology and Hepatology of Wisconsin, Milwaukee, USADepartment of Surgery of Medical College of Wisconsin, Milwaukee, USADepartment of Surgery of Medical College of Wisconsin, Milwaukee, USADepartment of Surgery of Medical College of Wisconsin, Milwaukee, USADivision of Gastroenterology and Hepatology of Wisconsin, Milwaukee, USADepartment of Surgery of Medical College of Wisconsin, Milwaukee, USADepartment of Pathology of Medical College of Wisconsin, Milwaukee, USADepartment of Visceral, Transplant, Thoracic and Vascular Surgery, University Hospital, Leipzig, GermanyDepartment of Pathology of Medical College of Wisconsin, Milwaukee, USADivision of Gastroenterology and Hepatology of Wisconsin, Milwaukee, USAINTRODUCTION: Wnt/β-catenin signaling activation has been reported only during the late steps of Barrett’s esophagus (BE) neoplastic progression, but not in BE metaplasia, based on the absence of nuclear β-catenin. However, β-catenin transcriptional activity has been recorded in absence of robust nuclear accumulation. Thus, we aimed to investigate the Wnt/β-catenin signaling in nondysplastic BE. METHODS: Esophageal tissues from healthy and BE patients without dysplasia were analyzed for Wnt target gene expression by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry. Esophageal squamous (EPC1-& EPC2-hTERT), BE metaplastic (CP-A), and adenocarcinoma (OE33) cell lines were characterized for Wnt activation by qRT-PCR, Western blot, and luciferase assay. Wnt activity regulation was examined by using recombinant Wnt3a and Dickkopf-1 (Dkk1) as well as Dkk1 short interfering RNA. RESULTS: Wnt target genes (AXIN2, c-MYC, Cyclin D1, Dkk1) and Wnt3a were significantly upregulated in nondysplastic BE compared with squamous mucosa. Elevated levels of dephosphorylated β-catenin were detected in nondysplastic BE. Nuclear active β-catenin and TOPflash activity were increased in CP-A and OE33 cells compared with squamous cells. Wnt3a-mediated β-catenin signaling activation was abolished by Dkk1 in CP-A cells. TOPFlash activity was elevated following Dkk1 silencing in CP-A but not in OE33 cells. Dysplastic and esophageal adenocarcinoma tissues demonstrated further Dkk1 and AXIN2 overexpression. CONCLUSIONS: Despite the absence of robust nuclear accumulation, β-catenin is transcriptionally active in nondysplastic BE. Dkk1 overexpression regulates β-catenin signaling in BE metaplastic but not in adenocarcinoma cells, suggesting that early perturbation of Dkk1-mediated signaling suppression may contribute to BE malignant transformation.http://www.sciencedirect.com/science/article/pii/S1476558615000871 |
| spellingShingle | Orestis Lyros Parvaneh Rafiee Linghui Nie Rituparna Medda Nebojsa Jovanovic Mary F. Otterson Behnaz Behmaram Ιnes Gockel Alexander Mackinnon Reza Shaker Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1 Neoplasia: An International Journal for Oncology Research |
| title | Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1 |
| title_full | Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1 |
| title_fullStr | Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1 |
| title_full_unstemmed | Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1 |
| title_short | Wnt/β-Catenin Signaling Activation beyond Robust Nuclear β-Catenin Accumulation in Nondysplastic Barrett’s Esophagus: Regulation via Dickkopf-1 |
| title_sort | wnt β catenin signaling activation beyond robust nuclear β catenin accumulation in nondysplastic barrett s esophagus regulation via dickkopf 1 |
| url | http://www.sciencedirect.com/science/article/pii/S1476558615000871 |
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