Epigenetic silencing of neurofilament genes promotes an aggressive phenotype in breast cancer

Neurofilament heavy polypeptide (NEFH) has recently been identified as a candidate DNA hypermethylated gene within the functional breast cancer hypermethylome. NEFH exists in a complex with neurofilament medium polypeptide (NEFM) and neurofilament light polypeptide (NEFL) to form neurofilaments, which are structural components of the cytoskeleton in mature neurons. Recent studies reported the deregulation of these proteins in several malignancies, suggesting that neurofilaments may have a role in other cell types as well. Using a comprehensive approach, we studied the epigenetic inactivation of neurofilament genes in breast cancer and the functional significance of this event. We report that DNA methylation-associated silencing of NEFH, NEFL, and NEFM in breast cancer is frequent, cancer-specific, and correlates with clinical features of disease progression. DNA methylation-mediated inactivation of these genes occurs also in multiple other cancer histologies including pancreas, gastric, and colon. Restoration of NEFH function, the major subunit of the neurofilament complex, reduces proliferation and growth of breast cancer cells and arrests them in Go/G1 phase of the cell cycle along with a reduction in migration and invasion. These findings suggest that DNA methylation-mediated silencing of the neurofilament genes NEFH, NEFM, and NEFL are frequent events that may contribute to the progression of breast cancer and possibly other malignancies.

[1]  Ying Zhuo,et al.  The role of NEFL in cell growth and invasion in head and neck squamous cell carcinoma cell lines. , 2014, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[2]  Mahmoud Abbas,et al.  Neurofilament Heavy polypeptide CpG island methylation associates with prognosis of renal cell carcinoma and prediction of antivascular endothelial growth factor therapy response , 2014, Cancer medicine.

[3]  Gianluca Bontempi,et al.  A comprehensive overview of Infinium HumanMethylation450 data processing , 2013, Briefings Bioinform..

[4]  S. Powers,et al.  Genome-wide methylation analysis and epigenetic unmasking identify tumor suppressor genes in hepatocellular carcinoma. , 2013, Gastroenterology.

[5]  R. Grimer,et al.  Functional epigenetic approach identifies frequently methylated genes in Ewing sarcoma , 2013, Epigenetics.

[6]  Ludmila V. Danilova,et al.  Frequent Inactivation of Cysteine Dioxygenase Type 1 Contributes to Survival of Breast Cancer Cells and Resistance to Anthracyclines , 2013, Clinical Cancer Research.

[7]  H. Putter,et al.  Chromatin remodeling of human subtelomeres and TERRA promoters upon cellular senescence , 2013, Epigenetics.

[8]  Pau Farré,et al.  Additional annotation enhances potential for biologically-relevant analysis of the Illumina Infinium HumanMethylation450 BeadChip array , 2013, Epigenetics & Chromatin.

[9]  L. Cope,et al.  Biomarkers for detection and prognosis of breast cancer identified by a functional hypermethylome screen , 2012, Epigenetics.

[10]  Xiao-Qing Li,et al.  NEFL mRNA Expression Level Is a Prognostic Factor for Early-Stage Breast Cancer Patients , 2012, PloS one.

[11]  S. Baylin,et al.  DNMT1 modulates gene expression without its catalytic activity partially through its interactions with histone-modifying enzymes , 2012, Nucleic acids research.

[12]  Martin J. Aryee,et al.  A DNA hypermethylation module for the stem/progenitor cell signature of cancer , 2011, Genome research.

[13]  C. Sotiriou,et al.  Evaluation of the Infinium Methylation 450K technology. , 2011, Epigenomics.

[14]  Peng Huang,et al.  Genome-wide methylation analysis identifies genes specific to breast cancer hormone receptor status and risk of recurrence. , 2011, Cancer research.

[15]  Peter A. Jones,et al.  A decade of exploring the cancer epigenome — biological and translational implications , 2011, Nature Reviews Cancer.

[16]  C. Ricketts,et al.  Genome-wide DNA methylation profiling of CpG islands in breast cancer identifies novel genes associated with tumorigenicity. , 2011, Cancer research.

[17]  Fang Fang,et al.  Breast Cancer Methylomes Establish an Epigenomic Foundation for Metastasis , 2011, Science Translational Medicine.

[18]  J. Herman,et al.  Genomic and Epigenomic Integration Identifies a Prognostic Signature in Colon Cancer , 2011, Clinical Cancer Research.

[19]  M. Fraga,et al.  DNA methylation epigenotypes in breast cancer molecular subtypes , 2010, Breast Cancer Research.

[20]  Martina Wiedau-Pazos,et al.  Integrative gene-tissue microarray-based approach for identification of human disease biomarkers: application to amyotrophic lateral sclerosis. , 2010, Human molecular genetics.

[21]  Juna Lee,et al.  Neurofilament Heavy Polypeptide Regulates the Akt-β-Catenin Pathway in Human Esophageal Squamous Cell Carcinoma , 2010, PloS one.

[22]  J. Herman,et al.  Methylation of TFPI2 in stool DNA: a potential novel biomarker for the detection of colorectal cancer. , 2009, Cancer research.

[23]  E. Vardar,et al.  beta-catenin gene mutation in invasive ductal breast cancer. , 2008, Journal of B.U.ON. : official journal of the Balkan Union of Oncology.

[24]  Gangning Liang,et al.  Frequent switching of Polycomb repressive marks and DNA hypermethylation in the PC3 prostate cancer cell line , 2008, Proceedings of the National Academy of Sciences.

[25]  T. Giordano,et al.  Genetic Changes of Wnt Pathway Genes Are Common Events in Metaplastic Carcinomas of the Breast , 2008, Clinical Cancer Research.

[26]  J. Herman,et al.  Epigenetic inactivation of the canonical Wnt antagonist SRY-box containing gene 17 in colorectal cancer. , 2008, Cancer research.

[27]  T. Mikkelsen,et al.  Genome-wide maps of chromatin state in pluripotent and lineage-committed cells , 2007, Nature.

[28]  Wei Chen,et al.  Comparing the DNA Hypermethylome with Gene Mutations in Human Colorectal Cancer , 2007, PLoS genetics.

[29]  S. Baylin,et al.  Stem Cell Chromatin Patterns: An Instructive Mechanism for DNA Hypermethylation? , 2007, Cell cycle.

[30]  Zohar Yakhini,et al.  Polycomb-mediated methylation on Lys27 of histone H3 pre-marks genes for de novo methylation in cancer , 2007, Nature Genetics.

[31]  Kelly M. McGarvey,et al.  A stem cell–like chromatin pattern may predispose tumor suppressor genes to DNA hypermethylation and heritable silencing , 2007, Nature Genetics.

[32]  A. Petzold.  Neurofilament phosphoforms: Surrogate markers for axonal injury, degeneration and loss , 2005, Journal of the Neurological Sciences.

[33]  S. Limborska,et al.  Analysis of heavy neurofilament subunit gene polymorphism in Russian patients with sporadic motor neuron disease (MND) , 2004, European Journal of Human Genetics.

[34]  J. Slingerland,et al.  Multiple Roles of the PI3K/PKB (Akt) Pathway in Cell Cycle Progression , 2003, Cell cycle.

[35]  J. Herman,et al.  Histone modifications and silencing prior to DNA methylation of a tumor suppressor gene. , 2003, Cancer cell.

[36]  J. Herman,et al.  Dependence of histone modifications and gene expression on DNA hypermethylation in cancer. , 2002, Cancer research.

[37]  O. Evgrafov,et al.  A new variant of Charcot-Marie-Tooth disease type 2 is probably the result of a mutation in the neurofilament-light gene. , 2000, American journal of human genetics.

[38]  J. Herman,et al.  Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[39]  W. Recant,et al.  Loss of heterozygosity from the short arm of chromosome 8 is associated with invasive behavior in breast cancer , 1996, Genes, chromosomes & cancer.

[40]  Y. Nakamura,et al.  Frequent loss of heterozygosity for loci on chromosome 8p in hepatocellular carcinoma, colorectal cancer, and lung cancer. , 1992, Cancer research.

[41]  Y. Nakamura,et al.  Allelotype of colorectal carcinomas. , 1989, Science.

[42]  P M Steinert,et al.  Molecular and cellular biology of intermediate filaments. , 1988, Annual review of biochemistry.

[43]  J W Griffin,et al.  Control of axonal caliber by neurofilament transport , 1984, The Journal of cell biology.

[44]  M. Shelanski,et al.  Intermediate filaments in nervous tissues , 1978, The Journal of cell biology.

[45]  R. Friede,et al.  Axon caliber related to neurofilaments and microtubules in sciatic nerve fibers of rats and mice , 1970, The Anatomical record.