Recurrent transcriptional loss of the PCDH17 tumor suppressor in laryngeal squamous cell carcinoma is partially mediated by aberrant promoter DNA methylation
Ewa Byzia
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorNatalia Soloch
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorMagdalena Bodnar
Department of Clinical Pathomorphology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Bydgoszcz, Poland
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorMarcin Szaumkessel
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorKatarzyna Kiwerska
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Department of Tumor Pathology, Greater Poland Cancer Center, Poznan, Poland
Search for more papers by this authorMagdalena Kostrzewska-Poczekaj
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorMalgorzata Jarmuz-Szymczak
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorLukasz Szylberg
Department of Clinical Pathomorphology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Bydgoszcz, Poland
Search for more papers by this authorMalgorzata Wierzbicka
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorAnna Bartochowska
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorEwelina Kalinowicz
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorReidar Grenman
Department of Otorhinolaryngology, -Head and Neck Surgery, Turku University Central Hospital and Turku University, Turku, Finland
Department of Medical Biochemistry, Turku University Central Hospital and Turku University, Turku, Finland
Search for more papers by this authorKrzysztof Szyfter
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorAndrzej Marszalek
Department of Tumor Pathology and Prophylaxis, Poznan University of Medical Sciences and Greater Poland Cancer Center, Poznan, Poland
Search for more papers by this authorCorresponding Author
Maciej Giefing
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Correspondence
Maciej Giefing, PhD, Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan/Poland.
Email: [email protected]
Search for more papers by this authorEwa Byzia
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorNatalia Soloch
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorMagdalena Bodnar
Department of Clinical Pathomorphology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Bydgoszcz, Poland
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorMarcin Szaumkessel
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorKatarzyna Kiwerska
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Department of Tumor Pathology, Greater Poland Cancer Center, Poznan, Poland
Search for more papers by this authorMagdalena Kostrzewska-Poczekaj
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorMalgorzata Jarmuz-Szymczak
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Department of Hematology and Bone Marrow Transplantation, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorLukasz Szylberg
Department of Clinical Pathomorphology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Torun, Bydgoszcz, Poland
Search for more papers by this authorMalgorzata Wierzbicka
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorAnna Bartochowska
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorEwelina Kalinowicz
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Search for more papers by this authorReidar Grenman
Department of Otorhinolaryngology, -Head and Neck Surgery, Turku University Central Hospital and Turku University, Turku, Finland
Department of Medical Biochemistry, Turku University Central Hospital and Turku University, Turku, Finland
Search for more papers by this authorKrzysztof Szyfter
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Search for more papers by this authorAndrzej Marszalek
Department of Tumor Pathology and Prophylaxis, Poznan University of Medical Sciences and Greater Poland Cancer Center, Poznan, Poland
Search for more papers by this authorCorresponding Author
Maciej Giefing
Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland
Department of Otolaryngology and Laryngological Oncology, Poznan University of Medical Sciences, Poznan, Poland
Correspondence
Maciej Giefing, PhD, Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan/Poland.
Email: [email protected]
Search for more papers by this authorAbstract
Protocadherins are cell-cell adhesion molecules encoded by a large family of genes. Recent reports demonstrate recurrent silencing of protocadherin genes in tumors and provide strong arguments for their tumor supresor functionality. Loss of protocadherins may contribute to cancer development not only by altering cell-cell adhesion, that is a hallmark of cancer, but also by enhancing proliferation and epithelial mesenchymal transition of cells via deregulation of the WNT signaling pathway. In this study we have further corroborated our previous findings on the involvement of PCDH17 in laryngeal squamous cell carcinoma (LSCC). We used bisulfite pyrosequencing to analyze a cohort of primary LSCC tumors for alterations in PCDH17 promoter DNA methylation as an alternative gene inactivation mechanism to the homozygous deletions reported earlier. Moreover, we analyzed primary LSCC samples by immunohistochemistry for PCDH17 protein loss. We identified recurrent elevation of PCDH17 promoter DNA methylation in 32/81 (40%) primary tumors (P < 0.001) and therein hypermethylation of 12 (15%) cases in contrast to no tumor controls (n = 24) that were all unmethylated. Importantly, DNA demethylation by decitabine has restored low level PCDH17 expression in LSCC cell lines. In conclusion, we provide a mechanistic explanation of recurrently observed PCDH17 silencing in LSCC by demonstrating the role of promoter methylation in this process. In light of these findings and recent reports showing that PCDH17 methylation is detectable in serum of cancer patients we suggest that testing PCDH17 DNA methylation might serve as a potential biomarker in LSCC.
Supporting Information
Additional Supporting Information may be found online in the supporting information tab for this article.
| Filename | Description |
|---|---|
| mc22808-sup-0001-SuppTab-S1.pdf549.4 KB |
Table S1. Clinical and pathological information of LSCC cell lines and primary samples investigated in this study. |
| mc22808-sup-0002-SuppTab-S2.pdf174.9 KB |
Table S2. Primers used for PCDH17 Sanger sequencing. |
| mc22808-sup-0003-SuppFig-S1.pdf276.6 KB |
Fig. S1. Bisulfite pyrosequencing based DNA methylation analysis of methylated DNA dilutions (0–100% concentrations with 10% increments). |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1 Frank M, Kemler R. Protocadherins. Curr Opin Cell Biol. 2002; 14: 557–562.
- 2 Keeler AB, Molumby MJ, Weiner JA. Protocadherins branch out: multiple roles in dendrite development. Cell Adh Migr. 2015; 9: 214–226.
- 3 Shan M, Su Y, Kang W, Gao R, Li X, Zhang G. Aberrant expression and functions of protocadherins in human malignant tumors. Tumour Biol. 2016; 37: 12969–12981.
- 4 El Hajj N, Dittrich M, Haaf T. Epigenetic dysregulation of protocadherins in human disease. Semin Cell Dev Biol. 2017; 69: 172–182.
- 5 Mah KM, Weiner JA. Regulation of Wnt signaling by protocadherins. Semin Cell Dev Biol. 2017; 69: 158–171.
- 6 DiMeo TA, Anderson K, Phadke P, et al. A novel lung metastasis signature links Wnt signaling with cancer cell self-renewal and epithelial-mesenchymal transition in basal-like breast cancer. Cancer Res. 2009; 69: 5364–5373.
- 7 Giefing M, Zemke N, Brauze D, et al. High resolution ArrayCGH and expression profiling identifies PTPRD and PCDH17/PCH68 as tumor suppressor gene candidates in laryngeal squamous cell carcinoma. Genes Chromosom Cancer. 2011; 50: 154–166.
- 8 Jarmuz M, Golusinski W, Grénman R, Szyfter K. Analysis of chromosome aberrations in cell lines derived from laryngeal cancer in relation to tumor progression. Eur Arch Otorhinolaryngol. 2002; 259: 269–273.
- 9 Jarvinen AK, Autio R, Haapa-Paananen S, et al. Identification of target genes in laryngeal squamous cell carcinoma by high-resolution copy number and gene expression microarray analyses. Oncogene. 2006; 25: 6997–7008.
- 10 Kiwerska K, Szaumkessel M, Paczkowska J, et al. Combined deletion and DNA methylation result in silencing of FAM107A gene in laryngeal tumors. Sci Rep. 2017; 7: 5386.
- 11 Chomczynski P. A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques. 1993; 15: 536–537.
- 12 Bodnar M, Szylberg L, Kazmierczak W, Marszalek A. Tumor progression driven by pathways activating matrix metalloproteinases and their inhibitors. J Oral Pathol Med. 2015; 44: 437–443.
- 13 Uhlen M, Oksvold P, Fagerberg L, et al. Towards a knowledge-based human protein atlas. Nat Biotechnol. 2010; 28: 1248–1250.
- 14 Remmele W, Stegner HE. Recommendation for uniform definition of an immunoreactive score (IRS) for immunohistochemical estrogen receptor detection (ER-ICA) in breast cancer tissue. Pathologe. 1987; 8: 138–140.
- 15 Szaumkessel M, Richter J, Giefing M, et al. Pyrosequencing-based DNA methylation profiling of Fanconi anemia/BRCA pathway genes in laryngeal squamous cell carcinoma. Int J Oncol. 2011; 39: 505–514.
- 16 Koressaar T, Remm M. Enhancements and modifications of primer design program Primer3. Bioinformatics. 2007; 23: 1289–1291.
- 17 Untergasser A, Cutcutache I, Koressaar T, et al. Primer3-new capabilities and interfaces. Nucleic Acids Res. 2012; 40: e115.
- 18 Kostrzewska-Poczekaj M, Giefing M, Jarmuz M, et al. Recurrent amplification in the 22q11 region in laryngeal squamous cell carcinoma results in overexpression of the CRKL but not the MAPK1 oncogene. Cancer Biomark. 2010-2011; 8: 11–19.
- 19 Haruki S, Imoto I, Kozaki K, et al. Frequent silencing of protocadherin 17, a candidate tumour suppressor for esophageal squamous cell carcinoma. Carcinogenesis. 2010; 31: 1027–1036.
- 20 Yin X, Xiang T, Mu J, et al. Protocadherin 17 functions as a tumor suppressor suppressing Wnt/β-catenin signaling and cell metastasis and is frequently methylated in breast cancer. Oncotarget. 2016; 7: 51720–51732.
- 21 Lin YL, Xie PG, Wang L, Ma JG. Aberrant methylation of protocadherin 17 and its clinical significance in patients with prostate cancer after radical prostatectomy. Med Sci Monit. 2014; 20: 1376–1382.
- 22 Lin YL, Wang YP, Li HZ, Zhang X. Aberrant promoter methylation of PCDH17 (Protocadherin 17) in serum and its clinical significance in renal cell carcinoma. Med Sci Monit. 2017; 23: 3318–3323.
- 23 Hu X, Sui X, Li L, et al. Protocadherin 17 acts as a tumour suppressor inducing tumour cell apoptosis and autophagy, and is frequently methylated in gastric and colorectal cancers. J Pathol. 2013; 229: 62–73.
- 24 Wang Y, Yu Y, Ye R, et al. An epigenetic biomarker combination of PCDH17 and POU4F2 detects bladder cancer accurately by methylation analyses of urine sediment DNA in Han Chinese. Oncotarget. 2016; 7: 2754–2764.
