The adenocarcinoma-associated antigen, AGR2, promotes tumor growth, cell migration, and cellular transformation.

The AGR2 gene encodes a secretory protein that is highly expressed in adenocarcinomas of the esophagus, pancreas, breast, and prostate. This study explores the effect of AGR2 expression with well-established in vitro and in vivo assays that screen for cellular transformation and tumor growth. AGR2 expression in SEG-1 esophageal adenocarcinoma cells was reduced with RNA interference. Cellular transformation was examined using NIH3T3 cells that express AGR2 after stable transfection. The cell lines were studied in vitro with assays for density-dependent and anchorage-independent growth, and in vivo as tumor xenografts in nude mice. SEG-1 cells with reduced AGR2 expression showed an 82% decrease in anchorage-independent colony growth and a 60% reduction in tumor xenograft size. In vitro assays of AGR2-expressing NIH3T3 cells displayed enhanced foci formation and anchorage-independent growth. In vivo, AGR2-expressing NIH3T3 cells established tumors in nude mice. Thus, AGR2 expression promotes tumor growth in esophageal adenocarcinoma cells and is able to transform NIH3T3 cells. Immunohistochemistry of the normal mouse intestine detected AGR2 expression in proliferating and differentiated intestinal cells of secretory lineage. AGR2 may be important for the growth and development of the intestine as well as esophageal adenocarcinomas.

[1]  P. Rudland,et al.  Significance of the metastasis-inducing protein AGR2 for outcome in hormonally treated breast cancer patients , 2006, British Journal of Cancer.

[2]  M. Bjerknes,et al.  Neurogenin 3 and the enteroendocrine cell lineage in the adult mouse small intestinal epithelium. , 2006, Developmental biology.

[3]  Irving L Weissman,et al.  Cancer stem cells--perspectives on current status and future directions: AACR Workshop on cancer stem cells. , 2006, Cancer research.

[4]  F. Aberger,et al.  Anterior specification of embryonic ectoderm: the role of the Xenopus cement gland-specific gene XAG-2 , 1998, Mechanisms of Development.

[5]  Masha Kocherginsky,et al.  Progression of Barrett's metaplasia to adenocarcinoma is associated with the suppression of the transcriptional programs of epidermal differentiation. , 2005, Cancer research.

[6]  C. Maley,et al.  Barrett's esophagus and its progression to adenocarcinoma. , 2006, Journal of the National Comprehensive Cancer Network : JNCCN.

[7]  R. Weigel,et al.  hAG-2, the human homologue of the Xenopus laevis cement gland gene XAG-2, is coexpressed with estrogen receptor in breast cancer cell lines. , 1998, Biochemical and biophysical research communications.

[8]  C. Leow,et al.  Hath 1 , Down-Regulated in Colon Adenocarcinomas , Inhibits Proliferation and Tumorigenesis of Colon Cancer Cells , 2004 .

[9]  M. Schenker,et al.  hAG-2 and hAG-3, human homologues of genes involved in differentiation, are associated with oestrogen receptor-positive breast tumours and interact with metastasis gene C4.4a and dystroglycan , 2003, British Journal of Cancer.

[10]  Christian Pilarsky,et al.  Expression profiling of microdissected matched prostate cancer samples reveals CD166/MEMD and CD24 as new prognostic markers for patient survival , 2005, The Journal of pathology.

[11]  M. Bjerknes,et al.  Intestinal epithelial stem cells and progenitors. , 2006, Methods in enzymology.

[12]  C. P. Leblond,et al.  Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. I. Columnar cell. , 1974, The American journal of anatomy.

[13]  R. Kuick,et al.  Differential expression of Hsp27 in normal oesophagus, Barrett's metaplasia and oesophageal adenocarcinomas , 1999, British Journal of Cancer.

[14]  J. Cheville,et al.  AGR2, an androgen‐inducible secretory protein overexpressed in prostate cancer , 2005, Genes, chromosomes & cancer.

[15]  M. Tomayko,et al.  Determination of subcutaneous tumor size in athymic (nude) mice , 2004, Cancer Chemotherapy and Pharmacology.

[16]  Patrick J. Paddison,et al.  Second-generation shRNA libraries covering the mouse and human genomes , 2005, Nature Genetics.

[17]  L. Montagnier,et al.  AGAR SUSPENSION CULTURE FOR THE SELECTIVE ASSAY OF CELLS TRANSFORMED BY POLYOMA VIRUS. , 1964, Virology.

[18]  Patrick O. Brown,et al.  Gene Expression Patterns in Pancreatic Tumors, Cells and Tissues , 2007, PloS one.

[19]  Dong Liu,et al.  Human homologue of cement gland protein, a novel metastasis inducer associated with breast carcinomas. , 2005, Cancer research.

[20]  Z. Werb,et al.  92-kD type IV collagenase mediates invasion of human cytotrophoblasts , 1991, The Journal of cell biology.

[21]  Thorsten Schmidt,et al.  Zinc finger protein GFI-1 has low oncogenic potential but cooperates strongly with pim and myc genes in T-cell lymphomagenesis , 1998, Oncogene.

[22]  M. Peifer,et al.  Wnt signaling in oncogenesis and embryogenesis--a look outside the nucleus. , 2000, Science.

[23]  H. Zoghbi,et al.  Gfi1 functions downstream of Math1 to control intestinal secretory cell subtype allocation and differentiation. , 2005, Genes & development.

[24]  J. Gingrich,et al.  Oxidative stress is the new stress , 2005, Nature Medicine.

[25]  H. Zoghbi,et al.  Intestine-specific ablation of mouse atonal homolog 1 (Math1) reveals a role in cellular homeostasis. , 2007, Gastroenterology.

[26]  Hans Clevers,et al.  Self-Renewal and Cancer of the Gut: Two Sides of a Coin , 2005, Science.

[27]  T. Hupp,et al.  The Barrett’s Antigen Anterior Gradient-2 Silences the p53 Transcriptional Response to DNA Damage* , 2004, Molecular & Cellular Proteomics.

[28]  Christine A Iacobuzio-Donahue,et al.  Exploration of global gene expression patterns in pancreatic adenocarcinoma using cDNA microarrays. , 2003, The American journal of pathology.

[29]  C. P. Leblond,et al.  Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. V. Unitarian Theory of the origin of the four epithelial cell types. , 1974, The American journal of anatomy.

[30]  M. Omary,et al.  Gene expression profiling reveals stromal genes expressed in common between Barrett's esophagus and adenocarcinoma. , 2006, Gastroenterology.

[31]  G. Kristiansen,et al.  Prognostic Relevance of AGR2 Expression in Breast Cancer , 2006, Clinical Cancer Research.

[32]  H. Zoghbi,et al.  Requirement of Math1 for Secretory Cell Lineage Commitment in the Mouse Intestine , 2001, Science.

[33]  N. Wright,et al.  Stem Cell Relationships and the Origin of Gastrointestinal Cancer , 2005, Oncology.

[34]  J. Abbruzzese,et al.  Developmental biology informs cancer: the emerging role of the hedgehog signaling pathway in upper gastrointestinal cancers. , 2003, Cancer cell.

[35]  C. Leow,et al.  Hath1, Down-Regulated in Colon Adenocarcinomas, Inhibits Proliferation and Tumorigenesis of Colon Cancer Cells , 2004, Cancer Research.

[36]  H. Okano,et al.  Musashi1: An Evolutionally Conserved Marker for CNS Progenitor Cells Including Neural Stem Cells , 2000, Developmental Neuroscience.

[37]  C. Der,et al.  Biological assays for cellular transformation. , 1994, Methods in enzymology.

[38]  Julian Lewis,et al.  Organizing cell renewal in the intestine: stem cells, signals and combinatorial control , 2006, Nature Reviews Genetics.

[39]  Isabelle Duluc,et al.  Neurogenin3 is differentially required for endocrine cell fate specification in the intestinal and gastric epithelium , 2002, The EMBO journal.

[40]  H Cheng,et al.  Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. IV. Paneth cells. , 1974, The American journal of anatomy.

[41]  C. Gilks,et al.  Growth Factor Independence-1 Is Expressed in Primary Human Neuroendocrine Lung Carcinomas and Mediates the Differentiation of Murine Pulmonary Neuroendocrine Cells , 2004, Cancer Research.

[42]  Hideyuki Okano,et al.  Identification of a putative intestinal stem cell and early lineage marker; musashi-1. , 2003, Differentiation; research in biological diversity.