Fascin overexpression promotes neoplastic progression in oral squamous cell carcinoma

BackgroundFascin is a globular actin cross-linking protein, which plays a major role in forming parallel actin bundles in cell protrusions and is found to be associated with tumor cell invasion and metastasis in various type of cancers including oral squamous cell carcinoma (OSCC). Previously, we have demonstrated that fascin regulates actin polymerization and thereby promotes cell motility in K8-depleted OSCC cells. In the present study we have investigated the role of fascin in tumor progression of OSCC.MethodsTo understand the role of fascin in OSCC development and/or progression, fascin was overexpressed along with vector control in OSCC derived cells AW13516. The phenotype was studied using wound healing, Boyden chamber, cell adhesion, Hanging drop, soft agar and tumorigenicity assays. Further, fascin expression was examined in human OSCC samples (N = 131) using immunohistochemistry and level of its expression was correlated with clinico-pathological parameters of the patients.ResultsFascin overexpression in OSCC derived cells led to significant increase in cell migration, cell invasion and MMP-2 activity. In addition these cells demonstrated increased levels of phosphorylated AKT, ERK1/2 and JNK1/2. Our in vitro results were consistent with correlative studies of fascin expression with the clinico-pathological parameters of the OSCC patients. Fascin expression in OSCC showed statistically significant correlation with increased tumor stage (P = 0.041), increased lymph node metastasis (P = 0.001), less differentiation (P = 0.005), increased recurrence (P = 0.038) and shorter survival (P = 0.004) of the patients.ConclusionIn conclusion, our results indicate that fascin promotes tumor progression and activates AKT and MAPK pathways in OSCC-derived cells. Further, our correlative studies of fascin expression in OSCC with clinico-pathological parameters of the patients indicate that fascin may prove to be useful in prognostication and treatment of OSCC.

[1]  L. Liotta,et al.  Metastatic potential correlates with enzymatic degradation of basement membrane collagen , 1980, Nature.

[2]  C. Verschraegen,et al.  Increased expression of fascin, motility associated protein, in cell cultures derived from ovarian cancer and in borderline and carcinomatous ovarian tumors , 2004, Clinical & Experimental Metastasis.

[3]  P. Rehak,et al.  Biologic significance of fascin expression in clear cell renal cell carcinoma: systematic analysis of primary and metastatic tumor tissues using a tissue microarray technique. , 2006, Urology.

[4]  S. Dalal,et al.  Novel function of keratins 5 and 14 in proliferation and differentiation of stratified epithelial cells , 2011, Molecular biology of the cell.

[5]  Judith Jones,et al.  Cell adhesion molecules, the extracellular matrix and oral squamous carcinoma. , 2007, International journal of oral and maxillofacial surgery.

[6]  C. Sawyers,et al.  The phosphatidylinositol 3-Kinase–AKT pathway in human cancer , 2002, Nature Reviews Cancer.

[7]  Artur Ferreira,et al.  Oral squamous cell carcinoma: review of prognostic and predictive factors. , 2006, Oral surgery, oral medicine, oral pathology, oral radiology, and endodontics.

[8]  D. Raben,et al.  Poor prognosis in patients with stage I and II oral tongue squamous cell carcinoma , 2008, Cancer.

[9]  D. Lev,et al.  Exposure of melanoma cells to dacarbazine results in enhanced tumor growth and metastasis in vivo. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[10]  A. Ingle,et al.  Suramin augments the antitumor and antimetastatic activity of pentoxifylline in B16F10 melanoma , 2007, International journal of cancer.

[11]  Y. Kudo,et al.  Invasion and Metastasis of Oral Cancer Cells Require Methylation of E-Cadherin and/or Degradation of Membranous β-Catenin , 2004, Clinical Cancer Research.

[12]  H. Ji,et al.  C-erbB-2/ HER-2 upregulates fascin, an actin-bundling protein associated with cell motility, in human breast cancer cell lines , 2000, Oncogene.

[13]  J. Woolgar,et al.  Histopathological prognosticators in oral and oropharyngeal squamous cell carcinoma. , 2006, Oral oncology.

[14]  P. McCrea,et al.  Fascin, an actin-bundling protein associated with cell motility, is upregulated in hormone receptor negative breast cancer , 1999 .

[15]  S. Dalal,et al.  Loss of keratins 8 and 18 leads to alterations in α6β4-integrin-mediated signalling and decreased neoplastic progression in an oral-tumour-derived cell line , 2011, Journal of Cell Science.

[16]  M. Al-Alwan,et al.  Fascin Is Involved in the Antigen Presentation Activity of Mature Dendritic Cells1 , 2001, The Journal of Immunology.

[17]  B. Gumbiner,et al.  Cell Adhesion: The Molecular Basis of Tissue Architecture and Morphogenesis , 1996, Cell.

[18]  Josephine C. Adams,et al.  Fascin, an actin-bundling protein, modulates colonic epithelial cell invasiveness and differentiation in vitro. , 2003, The American journal of pathology.

[19]  Josephine C. Adams,et al.  Fascins, and their roles in cell structure and function. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[20]  Josephine C. Adams,et al.  Dual actin-bundling and protein kinase C-binding activities of fascin regulate carcinoma cell migration downstream of Rac and contribute to metastasis. , 2007, Molecular biology of the cell.

[21]  E. Kieff,et al.  Epstein-Barr virus infection induces expression in B lymphocytes of a novel gene encoding an evolutionarily conserved 55-kilodalton actin-bundling protein , 1994, Journal of virology.

[22]  C. Cohan,et al.  Role of the actin bundling protein fascin in growth cone morphogenesis: localization in filopodia and lamellipodia. , 2001, Cell motility and the cytoskeleton.

[23]  Jeffrey N Myers,et al.  Disparity in pathologic and clinical lymph node staging in oral tongue carcinoma , 2003, Cancer.

[24]  S. Kannan,et al.  Loss of Keratin 8 Phosphorylation Leads to Increased Tumor Progression and Correlates with Clinico-Pathological Parameters of OSCC Patients , 2011, PloS one.

[25]  James S. Brown,et al.  Cervical lymph node metastasis in oral cancer: the importance of even microscopic extracapsular spread. , 2003, Oral oncology.

[26]  J. Shah,et al.  Changing concepts in the surgical management of the cervical node metastasis. , 2003, Oral oncology.

[27]  T. Mücke,et al.  Recurrence interval affects survival after local relapse of oral cancer. , 2009, Oral oncology.

[28]  J. Myers,et al.  Fascin over-expression is associated with aggressiveness of oral squamous cell carcinoma. , 2007, Cancer letters.

[29]  A. Jemal,et al.  Global cancer statistics , 2011, CA: a cancer journal for clinicians.

[30]  Jean Jakoncic,et al.  Migrastatin analogues target fascin to block tumour metastasis , 2011, Nature.

[31]  U. Pastorino,et al.  Independent prognostic value of fascin immunoreactivity in stage I nonsmall cell lung cancer , 2003, British Journal of Cancer.

[32]  E. Li,et al.  Role of fascin in the proliferation and invasiveness of esophageal carcinoma cells. , 2005, Biochemical and biophysical research communications.

[33]  A. Jemal,et al.  Global Cancer Statistics , 2011 .

[34]  S. Nieh,et al.  Expression of fascin in oral and oropharyngeal squamous cell carcinomas has prognostic significance – a tissue microarray study of 129 cases , 2007, Histopathology.

[35]  Tsutomu Nomura,et al.  Impact of lymph node metastasis on the pattern of failure and survival in oral carcinomas. , 2003, American journal of surgery.

[36]  J. Adams,et al.  Formation of stable microspikes containing actin and the 55 kDa actin bundling protein, fascin, is a consequence of cell adhesion to thrombospondin-1: implications for the anti-adhesive activities of thrombospondin-1. , 1995, Journal of cell science.

[37]  J. Westermarck,et al.  Regulation of matrix metalloproteinase expression in tumor invasion , 1999, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[38]  J. Cameron,et al.  Immunohistochemical validation of a novel epithelial and a novel stromal marker of pancreatic ductal adenocarcinoma identified by global expression microarrays: sea urchin fascin homolog and heat shock protein 47. , 2002, American journal of clinical pathology.

[39]  R. Hynes,et al.  Contact and adhesive specificities in the associations, migrations, and targeting of cells and axons , 1992, Cell.

[40]  C. Der,et al.  Targeting the Raf-MEK-ERK mitogen-activated protein kinase cascade for the treatment of cancer , 2007, Oncogene.

[41]  U. Pastorino,et al.  Independent value of fascin immunoreactivity for predicting lymph node metastases in typical and atypical pulmonary carcinoids. , 2003, Lung cancer.

[42]  P. Angadi,et al.  Aberrant β-catenin expression in the histologic differentiation of oral squamous cell carcinoma and verrucous carcinoma: an immunohistochemical study. , 2010, Journal of oral science.

[43]  Josephine C. Adams,et al.  Roles of fascin in human carcinoma motility and signaling: prospects for a novel biomarker? , 2005, The international journal of biochemistry & cell biology.

[44]  F. Matsumura,et al.  Cell-matrix adhesions differentially regulate fascin phosphorylation. , 1999, Molecular biology of the cell.

[45]  A. Ben-Ze'ev,et al.  Fascin, a novel target of beta-catenin-TCF signaling, is expressed at the invasive front of human colon cancer. , 2007, Cancer research.

[46]  J. Thiery.  Epithelial–mesenchymal transitions in tumour progression , 2002, Nature Reviews Cancer.

[47]  Zhong-ying Shen,et al.  Fascin is a potential biomarker for early-stage oesophageal squamous cell carcinoma , 2006, Journal of Clinical Pathology.

[48]  M. Vaidya,et al.  Implications of cytokeratin 8/18 filament formation in stratified epithelial cells: Induction of transformed phenotype , 2004, International journal of cancer.

[49]  S. Tsunoda,et al.  Prognostic Significance of Fascin Overexpression in Human Esophageal Squamous Cell Carcinoma , 2005, Clinical Cancer Research.

[50]  J. Rutka,et al.  The role of fascin in the migration and invasiveness of malignant glioma cells. , 2008, Neoplasia.

[51]  L. Kowalski,et al.  Clinicopathological and immunohistochemical evaluation of oral squamous cell carcinoma in patients with early local recurrence. , 2007, Oral oncology.

[52]  Josephine C. Adams,et al.  Roles of fascin in cell adhesion and motility. , 2004, Current opinion in cell biology.

[53]  E. Dowdle,et al.  Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. , 1980, Analytical biochemistry.

[54]  Josephine C. Adams,et al.  Prognostic significance of fascin expression in advanced colorectal cancer: an immunohistochemical study of colorectal adenomas and adenocarcinomas , 2006, BMC Cancer.

[55]  J. Ross,et al.  Actin‐binding protein fascin expression in skin neoplasia , 2002, Journal of cutaneous pathology.

[56]  R. C. Chang,et al.  Outcomes of oral squamous cell carcinoma in Taiwan after surgical therapy: factors affecting survival. , 2003, Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons.

[57]  S. Tsunoda,et al.  Effects of small interfering RNAs targeting fascin on human esophageal squamous cell carcinoma cell lines , 2010, Diagnostic pathology.

[58]  J. Myers,et al.  Metastasis of squamous cell carcinoma of the oral tongue , 2007, Cancer and Metastasis Reviews.

[59]  Maddy Parsons,et al.  Interaction of fascin and protein kinase Cα: a novel intersection in cell adhesion and motility , 2003, The EMBO journal.

[60]  E. Wagner,et al.  Signal integration by JNK and p38 MAPK pathways in cancer development , 2009, Nature Reviews Cancer.

[61]  G. Pelosi,et al.  Independent prognostic value of fascin immunoreactivity in stage III–IV colonic adenocarcinoma , 2007, British Journal of Cancer.

[62]  Gary G. Borisy,et al.  Role of fascin in filopodial protrusion , 2006, The Journal of cell biology.

[63]  J. Bryan,et al.  Fascin, an echinoid actin-bundling protein, is a homolog of the Drosophila singed gene product. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[64]  J. Ferlay,et al.  Global Cancer Statistics, 2002 , 2005, CA: a cancer journal for clinicians.

[65]  L. Machesky,et al.  Fascin: Invasive filopodia promoting metastasis. , 2010, Communicative & integrative biology.

[66]  J. Kim,et al.  Prognostic value of activated Akt expression in oral squamous cell carcinoma , 2005, Journal of Clinical Pathology.

[67]  P. Friedl.  Prespecification and plasticity: shifting mechanisms of cell migration. , 2004, Current opinion in cell biology.

[68]  Lei Cao,et al.  Altered expression of desmocollin 3, desmoglein 3, and β-catenin in oral squamous cell carcinoma: correlation with lymph node metastasis and cell proliferation , 2007, Virchows Archiv.

[69]  D. Cohen,et al.  PAR1b promotes cell-cell adhesion and inhibits dishevelled-mediated transformation of Madin-Darby canine kidney cells. , 2006, Molecular biology of the cell.

[70]  Y. Maehara,et al.  Fascin expression in progression and prognosis of hepatocellular carcinoma , 2009, Journal of surgical oncology.

[71]  F. Matsumura,et al.  Fascin, an actin-bundling protein, induces membrane protrusions and increases cell motility of epithelial cells. , 1998, Molecular biology of the cell.

[72]  Christine A Iacobuzio-Donahue,et al.  Highly expressed genes in pancreatic ductal adenocarcinomas: a comprehensive characterization and comparison of the transcription profiles obtained from three major technologies. , 2003, Cancer research.

[73]  J. S. Rao,et al.  Molecular mechanisms of glioma invasiveness: the role of proteases , 2003, Nature Reviews Cancer.