L1 Cell Adhesion Molecule Promotes Tumorigenicity and Metastatic Potential in Non–Small Cell Lung Cancer

Purpose: Non–small cell lung cancer (NSCLC) is a highly metastatic cancer with limited treatment options, thus requiring development of novel targeted therapies. Our group previously identified L1 cell adhesion molecule (L1CAM) expression as a member of a prognostic multigene expression signature for NSCLC patients. However, there is little information on the biologic function of L1CAM in lung cancer cells. This study investigates the functional and prognostic role of L1CAM in NSCLC. Experimental Design: Cox proportional hazards regression analysis was done on four independent published mRNA expression datasets of primary NSCLCs. L1CAM expression was suppressed by short-hairpin RNA (shRNA)–mediated silencing in human NSCLC cell lines. Effects were assessed by examining in vitro migration and invasion, in vivo tumorigenicity in mice, and metastatic potential using an orthotopic xenograft rat model of lung cancer. Results: L1CAM is an independent prognostic marker in resected NSCLC patients, with overexpression strongly associated with worse prognosis. L1CAM downregulation significantly decreased cell motility and invasiveness in lung cancer cells and reduced tumor formation and growth in mice. Cells with L1CAM downregulation were deficient in constitutive extracellular signal–regulated kinase (Erk) activation. Orthotopic studies showed that L1CAM suppression in highly metastatic lung cancer cells significantly decreases spread to distant organs, including bone and kidney. Conclusion: L1CAM is a novel prometastatic gene in NSCLC, and its downregulation may effectively suppress NSCLC tumor growth and metastasis. Targeted inhibition of L1CAM may be a novel therapy for NSCLC. Clin Cancer Res; 18(7); 1914–24. ©2012 AACR.

[1]  J. Crowley,et al.  The IASLC Lung Cancer Staging Project: Proposals for the Revision of the TNM Stage Groupings in the Forthcoming (Seventh) Edition of the TNM Classification of Malignant Tumours , 2007, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[2]  P. Altevogt,et al.  The cytoplasmic part of L1-CAM controls growth and gene expression in human tumors that is reversed by therapeutic antibodies , 2007, Oncogene.

[3]  A. Regev,et al.  SOX2 Is an Amplified Lineage Survival Oncogene in Lung and Esophageal Squamous Cell Carcinomas , 2009, Nature Genetics.

[4]  H. Kijima,et al.  L1 Cell adhesion molecule (L1CAM) expression at the cancer invasive front is a novel prognostic marker of pancreatic ductal adenocarcinoma , 2011, Journal of surgical oncology.

[5]  H. Asou,et al.  Molecular cloning of cDNA encoding the rat neural cell adhesion molecule L1 Two L1 isoforms in the cytoplasmic region are produced by differential splicing , 1991, FEBS letters.

[6]  Yi Zhang,et al.  Gene expression signatures for predicting prognosis of squamous cell and adenocarcinomas of the lung. , 2006, Cancer research.

[7]  M. Schachner,et al.  Overexpression of the cell adhesion molecule L1 is associated with metastasis in cutaneous malignant melanoma. , 2002, European journal of cancer.

[8]  E. Dejana,et al.  Adhesion molecule signalling: not always a sticky business , 2011, Nature Reviews Molecular Cell Biology.

[9]  P. Altevogt,et al.  Combined treatment of L1CAM antibodies and cytostatic drugs improve the therapeutic response of pancreatic and ovarian carcinoma. , 2012, Cancer letters.

[10]  I. Roninson,et al.  Cell adhesion molecule L1 disrupts E-cadherin-containing adherens junctions and increases scattering and motility of MCF7 breast carcinoma cells. , 2006, Cancer research.

[11]  H. Kamiguchi,et al.  Neural cell adhesion molecule L1: Signaling pathways and growth cone motility , 1997, Journal of neuroscience research.

[12]  M. Schachner,et al.  Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion. , 1984, The EMBO journal.

[13]  Igor Jurisica,et al.  Three-gene prognostic classifier for early-stage non small-cell lung cancer. , 2007, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[14]  A. Ben-Ze'ev,et al.  Nuclear factor-κB signaling and ezrin are essential for L1-mediated metastasis of colon cancer cells , 2010, Journal of Cell Science.

[15]  Igor Jurisica,et al.  Gene expression–based survival prediction in lung adenocarcinoma: a multi-site, blinded validation study , 2008, Nature Medicine.

[16]  C. Remé,et al.  The absence of c-fos prevents light-induced apoptotic cell death of photoreceptors in retinal degeneration in vivo , 1997, Nature Medicine.

[17]  D. Teplow,et al.  Neural adhesion molecule L1 as a member of the immunoglobulin superfamily with binding domains similar to fibronectin , 1988, Nature.

[18]  Y. Chae,et al.  L1 Expression as a Marker for Poor Prognosis, Tumor Progression, and Short Survival in Patients with Colorectal Cancer , 2007, Annals of Surgical Oncology.

[19]  P. Altevogt,et al.  L1 expression as a predictor of progression and survival in patients with uterine and ovarian carcinomas , 2003, The Lancet.

[20]  P. Altevogt,et al.  Efficient inhibition of intra-peritoneal tumor growth and dissemination of human ovarian carcinoma cells in nude mice by anti-L1-cell adhesion molecule monoclonal antibody treatment. , 2006, Cancer research.

[21]  G. Edelman,et al.  Neuron-glia cell adhesion molecule interacts with neurons and astroglia via different binding mechanisms , 1988, The Journal of cell biology.

[22]  O. Miettinen,et al.  Survival of Patients with Stage I Lung Cancer Detected on CT Screening , 2008 .

[23]  Marcin Skrzypski,et al.  An Immune Response Enriched 72-Gene Prognostic Profile for Early-Stage Non–Small-Cell Lung Cancer , 2009, Clinical Cancer Research.

[24]  R. Coleman.  Skeletal complications of malignancy , 1997, Cancer.

[25]  Robert Livingston,et al.  Vinorelbine plus cisplatin vs. observation in resected non-small-cell lung cancer. , 2005, The New England journal of medicine.

[26]  T. Brabletz,et al.  beta-catenin regulates the expression of the matrix metalloproteinase-7 in human colorectal cancer. , 1999, The American journal of pathology.

[27]  P. Altevogt,et al.  Ectodomain shedding of L1 adhesion molecule promotes cell migration by autocrine binding to integrins , 2001, The Journal of cell biology.

[28]  Igor Jurisica,et al.  Prognostic and predictive gene signature for adjuvant chemotherapy in resected non-small-cell lung cancer. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[29]  J Sijbers,et al.  L1 knockout mice show dilated ventricles, vermis hypoplasia and impaired exploration patterns. , 1998, Human molecular genetics.

[30]  V. Lemmon,et al.  RanBPM is an L1‐interacting protein that regulates L1‐mediated mitogen‐activated protein kinase activation , 2005, Journal of neurochemistry.

[31]  P. Altevogt,et al.  L1, a novel target of β-catenin signaling, transforms cells and is expressed at the invasive front of colon cancers , 2005, The Journal of cell biology.

[32]  P. Altevogt,et al.  L1 adhesion molecule on human lymphocytes and monocytes: expression and involvement in binding to αvβ3 integrin , 1996 .

[33]  M. McMahon,et al.  Extracellular Signal-regulated Kinase (ERK)-dependent Gene Expression Contributes to L1 Cell Adhesion Molecule-dependent Motility and Invasion* , 2004, Journal of Biological Chemistry.

[34]  Thomas Kirchner,et al.  β-Catenin Regulates the Expression of the Matrix Metalloproteinase-7 in Human Colorectal Cancer , 1999 .

[35]  P. Altevogt,et al.  The adhesion molecule L1 (CD171) promotes melanoma progression , 2006, International journal of cancer.

[36]  C. Ryu,et al.  L1 Cell Adhesion Molecule Is a Novel Therapeutic Target in Intrahepatic Cholangiocarcinoma , 2010, Clinical Cancer Research.

[37]  F. Feuerhake,et al.  Expression and prognostic value of L1-CAM in breast cancer. , 2009, Oncology reports.

[38]  R. Guillery,et al.  Errors in corticospinal axon guidance in mice lacking the neural cell adhesion molecule L1 , 1998, Current Biology.

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

[40]  I. Jurisica,et al.  Modeling of lung cancer by an orthotopically growing H460SM variant cell line reveals novel candidate genes for systemic metastasis , 2004, Oncogene.

[41]  M. Tsao,et al.  Modified gateway system for double shRNA expression and Cre/lox based gene expression , 2011, BMC biotechnology.

[42]  P. Altevogt,et al.  L1 adhesion molecule on human lymphocytes and monocytes: expression and involvement in binding to alpha v beta 3 integrin. , 1996, European journal of immunology.

[43]  G. Landreth,et al.  Activation of the MAPK Signal Cascade by the Neural Cell Adhesion Molecule L1 Requires L1 Internalization* , 1999, The Journal of Biological Chemistry.

[44]  Ming-Sound Tsao,et al.  K-ras mutations in non-small-cell lung carcinoma: a review. , 2006, Clinical lung cancer.

[45]  M. Johnston,et al.  Characterization of a highly metastatic, orthotopic lung cancer model in the nude rat , 1999, Clinical & Experimental Metastasis.

[46]  M. Schachner,et al.  Disruption of the mouse L1 gene leads to malformations of the nervous system , 1997, Nature Genetics.

[47]  P. Altevogt,et al.  L1 augments cell migration and tumor growth but not β3 integrin expression in ovarian carcinomas , 2005, International journal of cancer.

[48]  J. Pouysségur,et al.  Targeting the ERK signaling pathway in cancer therapy , 2006, Annals of medicine.

[49]  M. Tsao,et al.  Overview of molecular testing in non-small-cell lung cancer: mutational analysis, gene copy number, protein expression and other biomarkers of EGFR for the prediction of response to tyrosine kinase inhibitors , 2009, Oncogene.

[50]  Brian H. Dunford-Shore,et al.  Somatic mutations affect key pathways in lung adenocarcinoma , 2008, Nature.

[51]  D. Galileo,et al.  Soluble L1CAM promotes breast cancer cell adhesion and migration in vitro, but not invasion , 2010, Cancer Cell International.

[52]  P. Altevogt,et al.  L1CAM–integrin interaction induces constitutive NF-κB activation in pancreatic adenocarcinoma cells by enhancing IL-1β expression , 2010, Oncogene.