Genomic Profiling of Oral Squamous Cell Carcinoma by Array-Based Comparative Genomic Hybridization

We designed a study to investigate genetic relationships between primary tumors of oral squamous cell carcinoma (OSCC) and their lymph node metastases, and to identify genomic copy number aberrations (CNAs) related to lymph node metastasis. For this purpose, we collected a total of 42 tumor samples from 25 patients and analyzed their genomic profiles by array-based comparative genomic hybridization. We then compared the genetic profiles of metastatic primary tumors (MPTs) with their paired lymph node metastases (LNMs), and also those of LNMs with non-metastatic primary tumors (NMPTs). Firstly, we found that although there were some distinctive differences in the patterns of genomic profiles between MPTs and their paired LNMs, the paired samples shared similar genomic aberration patterns in each case. Unsupervised hierarchical clustering analysis grouped together 12 of the 15 MPT-LNM pairs. Furthermore, similarity scores between paired samples were significantly higher than those between non-paired samples. These results suggested that MPTs and their paired LNMs are composed predominantly of genetically clonal tumor cells, while minor populations with different CNAs may also exist in metastatic OSCCs. Secondly, to identify CNAs related to lymph node metastasis, we compared CNAs between grouped samples of MPTs and LNMs, but were unable to find any CNAs that were more common in LNMs. Finally, we hypothesized that subpopulations carrying metastasis-related CNAs might be present in both the MPT and LNM. Accordingly, we compared CNAs between NMPTs and LNMs, and found that gains of 7p, 8q and 17q were more common in the latter than in the former, suggesting that these CNAs may be involved in lymph node metastasis of OSCC. In conclusion, our data suggest that in OSCCs showing metastasis, the primary and metastatic tumors share similar genomic profiles, and that cells in the primary tumor may tend to metastasize after acquiring metastasis-associated CNAs.

[1]  Zubing Li,et al.  Efficacy of supraomohyoid neck dissection in patients with oral squamous cell carcinoma and negative neck. , 2006, American journal of surgery.

[2]  Yibin Kang,et al.  The Multifaceted Role of MTDH/AEG-1 in Cancer Progression , 2009, Clinical Cancer Research.

[3]  René Bernards,et al.  A progression puzzle. , 2002, Nature.

[4]  A. Jones.  Prognosis in mouth cancer: tumour factors. , 1994, European journal of cancer. Part B, Oral oncology.

[5]  S. Knuutila,et al.  Recurrent DNA copy number losses associated with metastasis of larynx carcinoma , 1999, Genes, chromosomes & cancer.

[6]  D. Brachman,et al.  Molecular biology of head and neck cancer. , 1994, Seminars in oncology.

[7]  Ash A. Alizadeh,et al.  Genome-wide analysis of DNA copy-number changes using cDNA microarrays , 1999, Nature Genetics.

[8]  Shu-Chun Lin,et al.  Array‐comparative genomic hybridization to detect genomewide changes in microdissected primary and metastatic oral squamous cell carcinomas , 2006, Molecular carcinogenesis.

[9]  A. Leonard,et al.  Correlation between prognosis and degree of lymph node involvement in carcinoma of the oral cavity. , 1978, American journal of surgery.

[10]  Pei Wang,et al.  Integrative analysis of DNA copy number and gene expression in metastatic oral squamous cell carcinoma identifies genes associated with poor survival , 2010, Molecular Cancer.

[11]  T. Helliwell.  Molecular markers of metastasis in squamous carcinomas , 2001, The Journal of pathology.

[12]  K. Polyak.  Is Breast Tumor Progression Really Linear? , 2008, Clinical Cancer Research.

[13]  W. Lam,et al.  Use of complete coverage array comparative genomic hybridization to define copy number alterations on chromosome 3p in oral squamous cell carcinomas. , 2003, Cancer research.

[14]  K. Kinzler,et al.  Cancer genes and the pathways they control , 2004, Nature Medicine.

[15]  H. Heng,et al.  The evolutionary mechanism of cancer , 2010, Journal of cellular biochemistry.

[16]  A. Mäkitie,et al.  Elective neck treatment versus observation in patients with T1/T2 N0 squamous cell carcinoma of oral tongue. , 2006, Oral oncology.

[17]  S. Tsugane,et al.  Tumor thickness predicts cervical metastasis in patients with stage I/II carcinoma of the tongue , 1998, Cancer.

[18]  E. Schröck,et al.  Genomic changes defining the genesis, progression, and malignancy potential in solid human tumors: A phenotype/genotype correlation , 1999, Genes, chromosomes & cancer.

[19]  C. Mathers,et al.  Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008 , 2010, International journal of cancer.

[20]  J. Aubin,et al.  Dual function of ERRα in breast cancer and bone metastasis formation: implication of VEGF and osteoprotegerin. , 2011, Cancer research.

[21]  Ash A. Alizadeh,et al.  Genome-wide analysis of DNA copy number variation in breast cancer using DNA microarrays , 1999, Nature Genetics.

[22]  William P Schiemann,et al.  Grb2 binding to Tyr284 in TbetaR-II is essential for mammary tumor growth and metastasis stimulated by TGF-beta. , 2008, Carcinogenesis.

[23]  Yoshimi Shoji,et al.  Combination effects of distinct cores in 11q13 amplification region on cervical lymph node metastasis of oral squamous cell carcinoma. , 2011, International journal of oncology.

[24]  P. Lichter,et al.  Recurrent copy number gain of transcription factor SOX2 and corresponding high protein expression in oral squamous cell carcinoma , 2010, Genes, chromosomes & cancer.

[25]  I. Petersen,et al.  Chromosomal alterations during metastasis formation of head and neck squamous cell carcinoma , 2002, Genes, chromosomes & cancer.

[26]  K. Sasaki,et al.  Intratumoral cytogenetic heterogeneity detected by comparative genomic hybridization and laser scanning cytometry in human gliomas. , 1998, Cancer research.

[27]  J. Andersen,et al.  Intratumor genomic heterogeneity in breast cancer with clonal divergence between primary carcinomas and lymph node metastases , 2007, Breast Cancer Research and Treatment.

[28]  G. Hampton,et al.  Cdc6 and cyclin E2 are PTEN-regulated genes associated with human prostate cancer metastasis. , 2009, Neoplasia.

[29]  R. Greenberg,et al.  Evaluation of size in prognosis of oral cancer , 1986, Cancer.

[30]  J. Segall,et al.  Twists and turns of invasion , 2012, Nature Cell Biology.

[31]  S. Hao,et al.  The role of supraomohyoid neck dissection in patients of oral cavity carcinoma. , 2002, Oral oncology.

[32]  J. Greenman,et al.  Can a genetic signature for metastatic head and neck squamous cell carcinoma be characterised by comparative genomic hybridisation? , 2004, British Journal of Cancer.

[33]  Haoyang Cai,et al.  arrayMap: A Reference Resource for Genomic Copy Number Imbalances in Human Malignancies , 2012, PloS one.

[34]  J. Troge,et al.  Tumour evolution inferred by single-cell sequencing , 2011, Nature.

[35]  Jonathan R. Clark,et al.  Established prognostic variables in NO oral carcinoma , 2006, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[36]  Jun Luo,et al.  Copy Number Analysis Indicates Monoclonal Origin of Lethal Metastatic Prostate Cancer , 2009, Nature Medicine.

[37]  Bradley P. Coe,et al.  Overexpression of LRP12, a gene contained within an 8q22 amplicon identified by high-resolution array CGH analysis of oral squamous cell carcinomas , 2004, Oncogene.

[38]  J. Peterse,et al.  Breast cancer metastasis: markers and models , 2005, Nature Reviews Cancer.

[39]  K. Okita,et al.  Interglandular cytogenetic heterogeneity detected by comparative genomic hybridization in pancreatic cancer. , 2002, Cancer research.

[40]  Carlos Gamallo,et al.  Elective neck dissection in early‐stage oral squamous cell carcinoma—does it influence recurrence and survival? , 2007, Head & neck.

[41]  G. Lockwood,et al.  Predictive value of tumor thickness for cervical lymph‐node involvement in squamous cell carcinoma of the oral cavity , 2009, Cancer.

[42]  C. Fisher,et al.  Tumour thickness predicts cervical nodal metastases and survival in early oral tongue cancer. , 2003, Oral oncology.

[43]  M. Carrozzo,et al.  Importance of tumor thickness and depth of invasion in nodal involvement and prognosis of oral squamous cell carcinoma: A review of the literature , 2005, Head & neck.

[44]  H. Heng,et al.  Genetic and epigenetic heterogeneity in cancer: A genome‐centric perspective , 2009, Journal of cellular physiology.

[45]  Chi-Hung Huang,et al.  RAC1 activation mediates Twist1-induced cancer cell migration , 2012, Nature Cell Biology.

[46]  T. Farias,et al.  Relevance of Skip Metastases for Squamous Cell Carcinoma of the Oral Tongue and the Floor of the Mouth , 2006, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[47]  L. Kuo,et al.  Differential gene expression signature between primary and metastatic head and neck squamous cell carcinoma , 2007, The Journal of pathology.

[48]  Bhuvanesh Singh,et al.  Genetic abnormalities associated with nodal metastasis in head and neck cancer , 2004, Head & neck.

[49]  Ian M. Wilson,et al.  Array CGH technologies and their applications to cancer genomes , 2005, Chromosome Research.

[50]  C. Perou,et al.  Molecular classification of head and neck squamous cell carcinomas using patterns of gene expression. , 2004, Cancer cell.

[51]  A. Schäffer,et al.  Clinicopathological and prognostic implications of genetic alterations in oral cancers. , 2011, Oncology letters.

[52]  Tzong-Ming Shieh,et al.  Genome‐wide profiling of oral squamous cell carcinoma , 2004, The Journal of pathology.

[53]  Roberto A. Lima,et al.  Elective Neck Dissection versus Observation in Stage I Squamous Cell Carcinomas of the Tongue and Floor of the Mouth , 2001, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[54]  C. Maley,et al.  Cancer is a disease of clonal evolution within the body1–3. This has profound clinical implications for neoplastic progression, cancer prevention and cancer therapy. Although the idea of cancer as an evolutionary problem , 2006 .

[55]  D. Sessions,et al.  The Influence of Lymph Node Metastasis in the Treatment of Squamous Cell Carcinoma of the Oral Cavity, Oropharynx, Larynx, and Hypopharynx: N0 Versus N+ , 2005, The Laryngoscope.

[56]  T. Furuya,et al.  Comparative genomic hybridization reveals genetic progression of oral squamous cell carcinoma from dysplasia via two different tumourigenic pathways , 2006, The Journal of pathology.

[57]  Bradley P. Coe,et al.  Novel regions of amplification on 8q distinct from the MYC locus and frequently altered in oral dysplasia and cancer , 2004, Genes, chromosomes & cancer.