Research Article

Elevated expression of histone demethylase PHF8 associates with adverse prognosis in patients of laryngeal and hypopharyngeal squamous cell carcinoma

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Lijun Liu

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Department of Otolaryngology Head & Neck Surgery, First Affiliated Hospital of South China University, Chuanshan Road 69, Hengyang, 421001, China

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Li She

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Haolei Tan

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Ming Wei

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Changhan Chen

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Zhongwu Su

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Donghai Huang

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Yongquan Tian

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Yuanzheng Qiu

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

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Yong Liu^‡

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

^‡Authors contributed equally

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Xin Zhang^‡

Department of Otolaryngology Head & Neck Surgery, Xiangya Hospital, Central South University, Xiangya Road 87, Changsha 410008, Hunan, China

Otolaryngology Major Disease Research Key Laboratory of Hunan Province, Changsha 410008, Hunan, China

^‡Authors contributed equally

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Published Online:15 Dec 2014https://doi.org/10.2217/epi.14.82

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Abstract

Aim: Overexpression of histone demethylase PHF8 has been reported to function as an oncoprotein in many cancers; however, the implications of PHF8 involvement in laryngeal and hypopharyngeal squamous cell carcinoma (LHSCC) remain unclear. This study aims to explore the expression of PHF8 and its clinical significance in LHSCC. Materials & methods: Western blotting and immunohistochemistry were performed to evaluate PHF8 protein expression in fresh and archived LHSCC samples. Global expressions of H3K27 and H3K9 methylation were analyzed in a cell line with PHF8 siRNA treatment. Results & conclusion: In our study, PHF8 was upregulated in fresh LHSCC tissues. Immunohistochemical staining revealed that the expression of PHF8 was positively associated with T classification, clinical stage, primary tumor position and tumor relapse. Survival analysis demonstrated that high PHF8 expression was significantly associated with shorter overall survival and disease-free survival. Moreover, PHF8 regulates the levels of H3K9me2 and H3K27me2 in LHSCC. Taken together, PHF8 might be a novel prognostic marker for this disease.

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Papers of special note have been highlighted as: • of interest; •• of considerable interest

References

1 Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J. Clin. 64(1), 9–29 (2014).Crossref, Medline, Google Scholar
2 Pisani P, Bray F, Parkin DM. Estimates of the world-wide prevalence of cancer for 25 sites in the adult population. Int. J. Cancer 97(1), 72–81 (2002).Crossref, Medline, CAS, Google Scholar
3 Argiris A, Karamouzis MV, Raben D, Ferris RL. Head and neck cancer. Lancet 371(9625), 1695–1709 (2008).Crossref, Medline, CAS, Google Scholar
4 Sturgis EM. The International Head and Neck Cancer Epidemiology Consortium. Head Neck 33(1), 141 (2011).Crossref, Medline, Google Scholar
5 Elsheikh MN, Rinaldo A, Hamakawa H et al. Importance of molecular analysis in detecting cervical lymph node metastasis in head and neck squamous cell carcinoma. Head Neck 28(9), 842–849 (2006).Crossref, Medline, Google Scholar
6 Russo N, Wang X, Liu M et al. A novel approach to biomarker discovery in head and neck cancer using an autoantibody signature. Oncogene 32(42), 5026–5037 (2013).Crossref, Medline, CAS, Google Scholar
7 Rozov SM, Berdnikov VA, Gorel FL, Lavrent'eva MV, Solonenko LP. [Structural characteristics of lysine-rich histone from the sperm of a mollusk Anodonta piscinalis]. Mol. Biol. (Mosk.) 18(6), 1497–1508 (1984).Medline, CAS, Google Scholar
8 Cheng CS, Rai K, Garber M et al. Semiconductor-based DNA sequencing of histone modification states. Nat. Commun. 4, 2672 (2013).Crossref, Medline, Google Scholar
9 Fischle W. Molecular mechanisms of histone modification function. Biochim. Biophys. Acta 1839(8), 621–622 (2014).Crossref, Medline, CAS, Google Scholar
10 Zhang Y, Reinberg D. Transcription regulation by histone methylation: interplay between different covalent modifications of the core histone tails. Genes Dev. 15(18), 2343–2360 (2001).Crossref, Medline, CAS, Google Scholar
11 Bannister AJ, Kouzarides T. Reversing histone methylation. Nature 436(7054), 1103–1106 (2005).Crossref, Medline, CAS, Google Scholar
12 Carr SM, Munro S, Zalmas LP et al. Lysine methylation-dependent binding of 53BP1 to the pRb tumor suppressor. Proc. Natl Acad. Sci. USA 111(31), 11341–11346 (2014).Crossref, Medline, CAS, Google Scholar
13 Barski A, Cuddapah S, Cui KR et al. High-resolution profiling of histone methylations in the human genome. Cell 129(4), 823–837 (2007).Crossref, Medline, CAS, Google Scholar
14 Bannister AJ, Zegerman P, Partridge JF et al. Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain. Nature 410(6824), 120–124 (2001).Crossref, Medline, CAS, Google Scholar
15 Lachner M, O'Carroll N, Rea S, Mechtler K, Jenuwein T. Methylation of histone H3 lysine 9 creates a binding site for HP1 proteins. Nature 410(6824), 116–120 (2001).Crossref, Medline, CAS, Google Scholar
16 Czermin B, Melfi R, Mccabe D, Seitz V, Imhof A, Pirrotta V. Drosophila enhancer of Zeste/ESC complexes have a histone H3 methyltransferase activity that marks chromosomal polycomb sites. Cell 111(2), 185–196 (2002).Crossref, Medline, CAS, Google Scholar
17 Yamane K, Tateishi K, Klose RJ et al. PLU-1 is an H3K4 demethylase involved in transcriptional repression and breast cancer cell proliferation. Mol. Cell 25(6), 801–812 Crossref, Medline, Google Scholar
18 Liu G, Bollig-Fischer A, Kreike B et al. Genomic amplification and oncogenic properties of the GASC1 histone demethylase gene in breast cancer. Oncogene 28(50), 4491–4500 (2009).Crossref, Medline, CAS, Google Scholar
19 Biron VL, Mohamed A, Hendzel MJ, Underhill DA, Seikaly H. Epigenetic differences between human papillomavirus-positive and -negative oropharyngeal squamous cell carcinomas. J. Otolaryngol. Head N. 41, S65–S70 (2012).Medline, Google Scholar
20 Gannon OM, De Long LM, Endo-Munoz L, Hazar-Rethinam M, Saunders NA. Dysregulation of the repressive H3K27 trimethylation mark in head and neck squamous cell carcinoma contributes to dysregulated squamous differentiation. Clin. Cancer Res. 19(2), 428–441 (2013).Crossref, Medline, CAS, Google Scholar
21 Bragado P, Estrada Y, Sosa MS et al. Analysis of marker-defined HNSCC subpopulations reveals a dynamic regulation of tumor initiating properties. PLoS ONE 7(1), e29974 (2012).Crossref, Medline, CAS, Google Scholar
22 Laytragoon-Lewin N, Castro J, Elmberger G et al. DNA methylation in tumour and normal mucosal tissue of head and neck squamous cell carcinoma (HNSCC) patients – new diagnostic approaches and treatment. Eur. J. Cancer 47, S545–S545 (2011).Crossref, Medline, Google Scholar
23 Feng WJ, Yonezawa M, Ye J, Jenuwein T, Grummt I. PHF8 activates transcription of rRNA genes through H3K4me3 binding and H3K9me1/2 demethylation. Nat. Struct. Mol. Biol. 17(4), 445–450 (2010).•• Reveals a functional interplay between H3K4 methylation and H3K9me1/2 demethylation by PHF8, which links dynamic histone methylation to rDNA transcription and neural disease.Crossref, Medline, CAS, Google Scholar
24 Yu L, Wang Y, Huang S et al. Structural insights into a novel histone demethylase PHF8. Cell Res. 20(2), 166–173 (2010).•• Sheds light on the molecular mechanism underlying PHF8-associated developmental and neurological diseases.Crossref, Medline, CAS, Google Scholar
25 Qi HH, Sarkissian M, Hu GQ et al. Histone H4K20/H3K9 demethylase PHF8 regulates zebrafish brain and craniofacial development. Nature 466(7305), 503–507 (2010).Crossref, Medline, CAS, Google Scholar
26 Pasini D, Hansen KH, Christensen J, Agger K, Cloos PaC, Helin K. Coordinated regulation of transcriptional repression by the RBP2 H3K4 demethylase and polycomb-repressive complex 2. Genes Dev. 22(10), 1345–1355 (2008).Crossref, Medline, CAS, Google Scholar
27 Shen YZ, Pan XF, Zhao H. The histone demethylase PHF8 is an oncogenic protein in human non-small cell lung cancer. Biochem. Biophys. Res. Commun. 451(1), 119–125 (2014).Crossref, Medline, CAS, Google Scholar
28 Bjorkman M, Ostling P, Harma V et al. Systematic knockdown of epigenetic enzymes identifies a novel histone demethylase PHF8 overexpressed in prostate cancer with an impact on cell proliferation, migration and invasion. Oncogene 31(29), 3444–3456 (2012).• First discovery that PHF8 is activated in a subset of prostate cancers and promotes cancer progression.Crossref, Medline, CAS, Google Scholar
29 Sun XJ, Qiu JJ, Zhu ST et al. Oncogenic features of PHF8 histone demethylase in esophageal squamous cell carcinoma. PLoS ONE 8(10), e77353 (2013).Crossref, Medline, CAS, Google Scholar
30 Lim HJ, Dimova NV, Tan MK, Sigoillot FD, King RW, Shi Y. The G2/M regulator histone demethylase PHF8 is targeted for degradation by the anaphase-promoting complex containing CDC20. Mol. Cell. Biol. 33(21), 4166–4180 (2013).Crossref, Medline, CAS, Google Scholar
31 Wang J, Lin XQ, Wang S et al. PHF8 and REST/NRSF co-occupy gene promoters to regulate proximal gene expression. Sci. Rep. 4, 5008 (2014).Crossref, Medline, CAS, Google Scholar
32 Asensio-Juan E, Gallego C, Martinez-Balbas MA. The histone demethylase PHF8 is essential for cytoskeleton dynamics. Nucleic Acids Res. 40(19), 9429–9440 (2012).Crossref, Medline, CAS, Google Scholar
33 Liu Y, Xie CL, Zhang X et al. Elevated expression of HMGB1 in squamous-cell carcinoma of the head and neck and its clinical significance. Eur. J. Cancer 46(16), 3007–3015 (2010).Crossref, Medline, CAS, Google Scholar
34 Zhu G, Cai G, Liu Y et al. Quantitative iTRAQ LC-MS/MS proteomics reveals transcription factor crosstalk and regulatory networks in hypopharyngeal squamous cell carcinoma. J. Cancer 5(7), 525–536 (2014).Crossref, Medline, CAS, Google Scholar
35 Filleron T, Laplanche A, Boher JM, Kramar A. An R function to non-parametric and piecewise analysis of competing risks survival data. Comput. Methods Programs Biomed. 100(1), 24–38 (2010).Crossref, Medline, Google Scholar
36 R Foundation for Statistical Computing. R: a language and environment for statistical computing. www.R-project.orgGoogle Scholar
37 Liu W, Tanasa B, Tyurina OV et al. PHF8 mediates histone H4 lysine 20 demethylation events involved in cell cycle progression. Nature 466(7305), 508–512 (2010).Crossref, Medline, CAS, Google Scholar
38 Yatim A, Benne C, Sobhian B et al. NOTCH1 nuclear interactome reveals key regulators of its transcriptional activity and oncogenic function. Mol. Cell 48(3), 445–458 (2012).Crossref, Medline, CAS, Google Scholar
39 Fortschegger K, De Graaf P, Outchkourov NS, Van Schaik FM, Timmers HT, Shiekhattar R. PHF8 targets histone methylation and RNA polymerase II to activate transcription. Mol. Cell. Biol. 30(13), 3286–3298 (2010).Crossref, Medline, CAS, Google Scholar
40 Wysocka J, Swigut T, Xiao H et al. A PHD finger of NURF couples histone H3 lysine 4 trimethylation with chromatin remodelling. Nature 442(7098), 86–90 (2006).Crossref, Medline, CAS, Google Scholar
41 Wend P, Fang L, Zhu Q et al. Wnt/beta-catenin signalling induces MLL to create epigenetic changes in salivary gland tumours. EMBO J. 32(14), 1977–1989 (2013).Crossref, Medline, CAS, Google Scholar
42 Sundar IK, Yao H, Rahman I. Oxidative stress and chromatin remodeling in chronic obstructive pulmonary disease and smoking-related diseases. Antioxid. Redox Signal. 18(15), 1956–1971 (2013).Crossref, Medline, CAS, Google Scholar
43 Johann DJ Jr, Mcguigan MD, Patel AR et al. Clinical proteomics and biomarker discovery. Ann. NY Acad. Sci. 1022, 295–305 (2004).Crossref, Medline, CAS, Google Scholar
44 Masood R, Hochstim C, Cervenka B et al. A novel orthotopic mouse model of head and neck cancer and lymph node metastasis. Oncogenesis 2, e68 (2013).Crossref, Medline, CAS, Google Scholar
45 Hojfeldt JW, Agger K, Helin K. Histone lysine demethylases as targets for anticancer therapy. Nat. Rev. Drug Discov. 12(12), 917–930 (2013).Crossref, Medline, CAS, Google Scholar

Vol. 7, No. 2

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Published online 15 December 2014

Published in print April 2015

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Acknowledgements

U Makendra (Emory University) and J Shi (School of Public Health, Central South University) are acknowledged for their suggestions.

Financial & competing interests disclosure

This study was funded by grants from the National High Technology Research and Development Program of China (the Chinese 863 Program, no. 2012AA02A201), the National Natural Science Foundation of China (no. 81202128, 81372906, 81372426, 81172558, 81472696 and 81272974), the Research Fund for the Doctoral Program of Higher Education of China (no. 20120162120049) and the Freedom Explore Program of Central South University (no. 2012QNZT099). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

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