Suppressive roles of calreticulin in prostate cancer growth and metastasis.

Calreticulin is an essential, multifunctional Ca(2+)-binding protein that participates in the regulation of intracellular Ca(2+) homeostasis, cell adhesion, and chaperoning. Calreticulin is abundantly expressed and regulated by androgens in prostate epithelial cells. Given the importance of both calreticulin in multiple essential cellular activities and androgens in prostate cancer, we investigated the possibility of a role for calreticulin in prostate cancer progression. Immunohistochemistry revealed the down-regulation of calreticulin in a subset of human prostate cancer specimens. Prostate cancer cells overexpressing exogenous calreticulin produced fewer colonies in both monolayer culture and soft agar. Furthermore, calreticulin overexpression also inhibited tumor growth in the orthotopic PC3 xenograft tumor model and macroscopic lung metastasis in the rat Dunning AT3.1 prostate tumor model. To address the potential mechanism of calreticulin suppression of prostate cancer, we generated calreticulin mutants with different functional domains deleted. The calreticulin mutants containing the P-domain, which binds to other endoplasmic reticulum chaperone proteins, were sufficient for the suppression of PC3 growth in colony formation assays. Overall, our data support the hypothesis that calreticulin inhibits growth and/or metastasis of prostate cancer cells and that this suppression requires the P-domain.

[1]  É. Szabó,et al.  Kinase-dependent adhesion to fibronectin: regulation by calreticulin. , 2008, Experimental cell research.

[2]  A. Jemal,et al.  Cancer Statistics, 2008 , 2008, CA: a cancer journal for clinicians.

[3]  Rajiv Dhir,et al.  Tissue microarrays: an overview. , 2008, Methods in molecular biology.

[4]  M. Opas,et al.  Dissecting focal adhesions in cells differentially expressing calreticulin: a microscopy study , 2007, Biology of the cell.

[5]  C. McCulloch,et al.  Calreticulin Affects Fibronectin-based Cell-Substratum Adhesion via the Regulation of c-Src Activity* , 2007, Journal of Biological Chemistry.

[6]  Zhou Wang,et al.  The Hsp90 inhibitor, 17‐AAG, prevents the ligand‐independent nuclear localization of androgen receptor in refractory prostate cancer cells , 2007, The Prostate.

[7]  K. Mabuchi,et al.  The calcium- and zinc-responsive regions of calreticulin reside strictly in the N-/C-domain. , 2006, Biochimica et biophysica acta.

[8]  S. Eggener,et al.  Enhancement of intermittent androgen ablation by “off‐cycle” maintenance with finasteride in LNCaP prostate cancer xenograft model , 2006, The Prostate.

[9]  Yinan Zhang,et al.  Assembly of MHC class I molecules within the endoplasmic reticulum , 2006, Immunologic research.

[10]  Maurizio Molinari,et al.  The Use of Calnexin and Calreticulin by Cellular and Viral Glycoproteins* , 2005, Journal of Biological Chemistry.

[11]  M. Michalak,et al.  Calreticulin, a Ca2+-binding chaperone of the endoplasmic reticulum. , 2005, The international journal of biochemistry & cell biology.

[12]  T. Kawaguchi.  Cancer metastasis: characterization and identification of the behavior of metastatic tumor cells and the cell adhesion molecules, including carbohydrates. , 2005, Current drug targets. Cardiovascular & haematological disorders.

[13]  Konstantinos N. Syrigos,et al.  cancer xenograft model , 2004 .

[14]  Zhou Wang,et al.  Suppression of prostate tumor growth by U19, a novel testosterone-regulated apoptosis inducer. , 2003, Cancer research.

[15]  D. Mercola,et al.  Inhibition of Egr-1 expression reverses transformation of prostate cancer cells in vitro and in vivo , 2003, Oncogene.

[16]  K. Krause,et al.  Functional specialization of calreticulin domains , 2001, The Journal of cell biology.

[17]  M Opas,et al.  Calreticulin: one protein, one gene, many functions. , 1999, The Biochemical journal.

[18]  A. M. Carmo,et al.  Calreticulin Is Expressed on the Cell Surface of Activated Human Peripheral Blood T Lymphocytes in Association with Major Histocompatibility Complex Class I Molecules* , 1999, The Journal of Biological Chemistry.

[19]  C. Lo,et al.  Calreticulin Affects Focal Contact-dependent but Not Close Contact-dependent Cell-substratum Adhesion* , 1999, The Journal of Biological Chemistry.

[20]  Z. Wang,et al.  Calreticulin expression is associated with androgen regulation of the sensitivity to calcium ionophore-induced apoptosis in LNCaP prostate cancer cells. , 1999, Cancer research.

[21]  K. Krause,et al.  Calreticulin Is Essential for Cardiac Development , 1999, The Journal of cell biology.

[22]  E. B. Pewitt,et al.  Calreticulin: An Intracellular Ca++-Binding Protein Abundantly Expressed and Regulated by Androgen in Prostatic Epithelial Cells* * This work was supported by Boehringer Ingelheim International GmbH, American Cancer Society, Illinois Division Grant 95-58, the Robert H. Lurie Cancer Center Grant 200, , 1998, Endocrinology.

[23]  M. L. Le Beau,et al.  Identification of a novel metastasis-suppressor region on human chromosome 12. , 1998, Cancer research.

[24]  David Y. Thomas,et al.  Enhanced Catalysis of Ribonuclease B Folding by the Interaction of Calnexin or Calreticulin with ERp57* , 1998, The Journal of Biological Chemistry.

[25]  M. Michalak,et al.  Oligosaccharide binding characteristics of the molecular chaperones calnexin and calreticulin. , 1998, Biochemistry.

[26]  E. B. Pewitt,et al.  Calreticulin: an intracellular Ca++-binding protein abundantly expressed and regulated by androgen in prostatic epithelial cells. , 1998, Endocrinology.

[27]  X. Cai,et al.  Genes regulated by androgen in the rat ventral prostate. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[28]  F. Schröder,et al.  Orthotopic implantation of human prostate cancer cell lines: A clinically relevant animal model for metastatic prostate cancer , 1997, The Prostate.

[29]  S. Grinstein,et al.  Calreticulin is essential for integrin-mediated calcium signalling and cell adhesion , 1997, Nature.

[30]  J. Isaacs,et al.  CD44 is a metastasis suppressor gene for prostatic cancer located on human chromosome 11p13. , 1997, Cancer research.

[31]  M. Michalak,et al.  Calreticulin modulates cell adhesiveness via regulation of vinculin expression , 1996, The Journal of cell biology.

[32]  M Opas,et al.  Endoplasmic Reticulum Form of Calreticulin Modulates Glucocorticoid-sensitive Gene Expression* , 1996, The Journal of Biological Chemistry.

[33]  K. Krause,et al.  Overexpression of Calreticulin Increases Intracellular Ca Storage and Decreases Store-operated Ca Influx (*) , 1996, The Journal of Biological Chemistry.

[34]  S. Baksh,et al.  Identification of the Zn2+ binding region in calreticulin , 1995, FEBS letters.

[35]  S. Dedhar,et al.  Inducible interaction of integrin alpha 2 beta 1 with calreticulin. Dependence on the activation state of the integrin. , 1995, The Journal of biological chemistry.

[36]  P. Camacho,et al.  Calreticulin inhibits repetitive intracellular Ca2+ waves , 1995, Cell.

[37]  A. Helenius,et al.  Transient, lectin-like association of calreticulin with folding intermediates of cellular and viral glycoproteins. , 1995, Molecular biology of the cell.

[38]  E. Clementi,et al.  Overexpression of calreticulin increases the Ca2+ capacity of rapidly exchanging Ca2+ stores and reveals aspects of their lumenal microenvironment and function , 1995, The Journal of cell biology.

[39]  J. D. Capra,et al.  The unveiling of calreticulin--a clinically relevant tour of modern cell biology. , 1995, Journal of investigative medicine : the official publication of the American Federation for Clinical Research.

[40]  Qiang Zhu,et al.  Cell Surface Calreticulin Is a Putative Mannoside Lectin Which Triggers Mouse Melanoma Cell Spreading (*) , 1995, The Journal of Biological Chemistry.

[41]  W. Nauseef,et al.  Calreticulin Functions as a Molecular Chaperone in the Biosynthesis of Myeloperoxidase (*) , 1995, The Journal of Biological Chemistry.

[42]  E. Simons,et al.  Decreasing calreticulin expression lowers the Ca2+ response to bradykinin and increases sensitivity to ionomycin in NG-108-15 cells. , 1994, The Journal of biological chemistry.

[43]  W. Diekmann,et al.  Retention and retrieval: both mechanisms cooperate to maintain calreticulin in the endoplasmic reticulum. , 1994, Journal of cell science.

[44]  S. Dedhar.  Novel functions for calreticulin: interaction with integrins and modulation of gene expression? , 1994, Trends in biochemical sciences.

[45]  S. Dedhar,et al.  Cell attachment to extracellular matrix substrates is inhibited upon downregulation of expression of calreticulin, an intracellular integrin alpha-subunit-binding protein. , 1994, Journal of cell science.

[46]  M. Shago,et al.  Inhibition of nuclear hormone receptor activity by calreticulin , 1994, Nature.

[47]  K. Burns,et al.  Modulation of gene expression by calreticulin binding to the glucocorticoid receptor , 1994, Nature.

[48]  E. Wilson,et al.  The androgen receptor: an overview. , 1994, Recent progress in hormone research.

[49]  C. Dinney,et al.  Metastatic model for human prostate cancer using orthotopic implantation in nude mice. , 1992, Journal of the National Cancer Institute.

[50]  R. Gittes.  Carcinoma of the prostate. , 1991, The New England journal of medicine.

[51]  S. Kukreja,et al.  Patterns of metastasis by the human prostate cancer cell line PC‐3 in athymic nude mice , 1989, The Prostate.

[52]  W. Isaacs,et al.  Establishment and characterization of seven dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers , 1987, The Prostate.

[53]  J. Lechner,et al.  Establishment and characterization of a human prostatic carcinoma cell line (PC-3). , 1979, Investigative urology.

[54]  J. Mulcahy,et al.  Carcinoma of the prostate. , 1978, The Journal of the Kentucky Medical Association.

[55]  Y. Oshika,et al.  P-glycoprotein-mediated acquired multidrug resistance of human lung cancer cells in vivo. , 1996, British Journal of Cancer.