Abstract
Wnt10b is a signaling protein regulating skin development and homeostasis, and the expression of Wnt10b is restricted to epidermal keratinocytes in embryonic and postnatal skin. Recent studies indicate an elevated expression of Wnt10b in skin tumors. However, how Wnt10b regulates skin tumorigenesis remains largely unknown. Here we report that continuous expression of Wnt10b mediates transformation of epidermal keratinocytes through activating genes involved in EGF/MAPK signaling pathways. We first established a prolonged Wnt10b overexpression system in JB6P− cells to represent the elevated Wnt10b expression level in skin keratinocytes. Through expression assays and observations under phase-contrast microscopy, prolonged expression of Wnt10b activated Wnt/β-catenin pathway and induced morphological changes of cells showing longer protrusions and multilayer growth, indicating early-stage cell transformation. Wnt10b also increased cellular proliferation and migration according to BrdU incorporation and cell mobility assays. Furthermore, multi-doses of AdWnt10b treatment to JB6P− cells induced colony formation, stronger invasive ability in transwell system, and anchorage-independent growth in agar gel. In molecular level, AdWnt10b treatment induced increased transcriptional expressions of Egf, downstream Mapk pathway factors, and MMPs. Administration of Wnt antagonist DKK1 blocked the tumor promotion process induced by Wnt10b. Taken together, these findings clearly demonstrate that Wnt10b promotes epidermal keratinocyte transformation through induced Egf pathway.
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Abbreviations
- AdGFP:
-
Control adenoviruses
- AdWnt10b:
-
Wnt10b adenoviruses
- AP-1:
-
Activator protein
- BCC:
-
Basal cell carcinoma
- DKK1:
-
Dickkopf 1
- ECM:
-
Extracellular matrix
- EGF:
-
Epidermal growth factor
- NF-KB:
-
Nuclear factor kappa B
- PAGE:
-
Polyacrylamide gel electrophoresis
- PBS:
-
Phosphate-buffered saline
- PVDF:
-
Polyvinylidene fluoride
- RT-PCR:
-
Reverse transcription-polymerase chain reaction
- SCC:
-
Squamous cell carcinomas
- SDS:
-
Sodium dodecyl sulfate
- TNF-a:
-
Tumor necrosis factor-a
- TPA:
-
12-O-tetradecanoylphorbol-13-acetate
- Wnt10b:
-
Wingless-type MMTV integration site family, member 10B
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Acknowledgments
This study was supported by Grants 81171515 from National Nature Science Foundation of China and CSTC; Innovation and Attracting Talents Program for College and University (‘111’ Project) (B06023), China; and the “BaYu” Scholars Project in Chongqing for Dr. Cheng-Ming Chuong. We thank Dr. Pin-Chi Tang (National Chung Hsing University) and Dr. Ya-Chen Liang (University of Southern California) for carefully revising the manuscript.
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M. Lei and X. Lai have contributed equally to this work.
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Lei, M., Lai, X., Bai, X. et al. Prolonged overexpression of Wnt10b induces epidermal keratinocyte transformation through activating EGF pathway. Histochem Cell Biol 144, 209–221 (2015). https://doi.org/10.1007/s00418-015-1330-6
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DOI: https://doi.org/10.1007/s00418-015-1330-6