Analysis of host- and tumor-derived proteinases using a custom dual species microarray reveals a protective role for stromal matrix metalloproteinase-12 in non-small cell lung cancer.

We used a customized Affymetrix protease microarray (Hu/Mu ProtIn chip) designed to distinguish human and mouse genes to analyze the expression of proteases and protease inhibitors in lung cancer. Using an orthotopic lung cancer model, we showed that murine matrix metalloproteinase (MMP)-12, MMP-13, and cathepsin K were up-regulated in tumor tissue compared with normal mouse lung. To determine the relevance of stromal proteases detected using this model system, we compared the results to an analysis of human lung adenocarcinoma specimens using the U133 Plus 2.0 Affymetrix microarray. MMP-12, MMP-13, and cathepsin K showed an increase in expression in human tumors compared with normal lung similar to that seen in the orthotopic model. Immunohistochemical analysis confirmed MMP-12 expression in the stroma of human lung tumor samples. To determine the biological relevance of stromal MMP-12, murine Lewis lung carcinoma cells were injected into the tail vein of syngeneic wild-type (WT) and MMP-12-null mice. MMP-12-null and WT mice developed equivalent numbers of lung tumors; however, there was a 2-fold increase in the number of tumors that reached >2 mm in diameter in MMP-12-null mice compared with WT controls. The increase in tumor size correlated with an increase in CD31-positive blood vessels and a decrease in circulating levels of the K1-K4 species of angiostatin. These results show a protective role for stromal MMP-12 in lung tumor growth. The use of the Hu/Mu ProtIn chip allows us to distinguish tumor- and host-derived proteases and guides the further analysis of the significance of these genes in tumor progression.

[1]  R. Shoemaker,et al.  Novel intrapulmonary model for orthotopic propagation of human lung cancers in athymic nude mice. , 1987, Cancer research.

[2]  J. Folkman,et al.  The role of angiogenesis in tumor growth. , 1992, Seminars in cancer biology.

[3]  T. Ley,et al.  Metalloelastase is required for macrophage-mediated proteolysis and matrix invasion in mice. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[4]  I. Fidler,et al.  Macrophage-Derived Metalloelastase Is Responsible for the Generation of Angiostatin in Lewis Lung Carcinoma , 1997, Cell.

[5]  T. Thompson,et al.  Requirement for matrix metalloproteinase-9 (gelatinase B) expression in metastasis by murine prostate carcinoma. , 1998, The American journal of pathology.

[6]  M. Mizumoto,et al.  Expression of human macrophage metalloelastase gene in hepatocellular carcinoma: Correlation with angiostatin generation and its clinical significance , 1998, Hepatology.

[7]  M. d’Ortho,et al.  Role of collagenase in mediating in vitro alveolar epithelial wound repair. , 1999, Journal of cell science.

[8]  S. Shapiro.  Diverse Roles of Macrophage Matrix Metalloproteinases in Tissue Destruction and Tumor Growth , 1999, Thrombosis and Haemostasis.

[9]  T. Welte,et al.  Expression of cathepsin K in lung epithelial cells. , 1999, American journal of respiratory cell and molecular biology.

[10]  R. Khokha,et al.  Differential expression of matrix metalloproteinases and their inhibitors in non‐small cell lung cancer , 2000, The Journal of pathology.

[11]  T. Welte,et al.  Cathepsin K expression in human lung. , 2000, Advances in experimental medicine and biology.

[12]  A. Pozzi,et al.  Elevated matrix metalloprotease and angiostatin levels in integrin alpha 1 knockout mice cause reduced tumor vascularization. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Hanahan,et al.  MMP-9 Supplied by Bone Marrow–Derived Cells Contributes to Skin Carcinogenesis , 2000, Cell.

[14]  Bonnie F. Sloane,et al.  Inhibitors of Papain-Like Cysteine Peptidases in Cancer , 2000 .

[15]  S. Arii,et al.  Human macrophage metalloelastase gene expression in colorectal carcinoma and its clinicopathologic significance , 2001, Cancer.

[16]  Elise C. Kohn,et al.  The microenvironment of the tumour–host interface , 2001, Nature.

[17]  A. Pozzi,et al.  Low plasma levels of matrix metalloproteinase 9 permit increased tumor angiogenesis , 2002, Oncogene.

[18]  S. Shapiro,et al.  Metalloelastase (MMP‐12) expression by tumour cells in squamous cell carcinoma of the vulva correlates with invasiveness, while that by macrophages predicts better outcome , 2002, The Journal of pathology.

[19]  Yu Shyr,et al.  A training-testing approach to the molecular classification of resected non-small cell lung cancer. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[20]  C. López-Otín,et al.  Loss of collagenase-2 confers increased skin tumor susceptibility to male mice , 2003, Nature Genetics.

[21]  R. Hynes,et al.  Physiological levels of tumstatin, a fragment of collagen IV alpha3 chain, are generated by MMP-9 proteolysis and suppress angiogenesis via alphaV beta3 integrin. , 2003, Cancer cell.

[22]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[23]  L. Matrisian,et al.  A Protective Role for Matrix Metalloproteinase-3 in Squamous Cell Carcinoma , 2004, Cancer Research.

[24]  B. Fingleton,et al.  Expansion of myeloid immune suppressor Gr+CD11b+ cells in tumor-bearing host directly promotes tumor angiogenesis. , 2004, Cancer cell.

[25]  Bonnie F. Sloane,et al.  Proteases, extracellular matrix, and cancer: a workshop of the path B study section. , 2004, The American journal of pathology.

[26]  T. Welte,et al.  Pivotal role of cathepsin K in lung fibrosis. , 2004, The American journal of pathology.

[27]  Günther Richter,et al.  Matrix metalloproteinase-12 expression correlates with local recurrence and metastatic disease in non-small cell lung cancer patients. , 2005, Clinical cancer research : an official journal of the American Association for Cancer Research.

[28]  Heath B Acuff,et al.  An Orthotopic Model of Lung Cancer to Analyze Primary and Metastatic NSCLC Growth in Integrin α1-Null Mice , 2005, Clinical & Experimental Metastasis.