title: Role of notch-1 and E-cadherin in the differential response to calcium in culturing normal versus malignant prostate cells.
abstract: A panel of expression markers was validated and used to document that, when radical prostatectomy specimens are cultured in low (i.e., <260 micromol/L)-calcium (Ca2+)-serum-free, growth factor-defined (SFD) medium, what grows out are not prostatic cancer cells but basally derived normal transit-amplifying prostatic epithelial cells. The selective outgrowth of the normal transit-amplifying versus prostatic cancer cells is due to the differential effect of low-Ca2+ medium on the structure of Notch-1 and E-cadherin signaling molecules. In low-Ca2+ medium, Notch-1 receptor is conformationally in a constitutively active, cell autonomous form not requiring reciprocal cell-cell (i.e., ligand) interaction for signaling. Such signaling is required for survival of transit-amplifying cells as shown by the death of transit-amplifying cells induced by treatment with a series of chemically distinct gamma-secretase inhibitors to prevent Notch-1 signaling. Conversely, in low-Ca2+ medium, E-cadherin is conformationally inactive preventing cell-cell homotypic interaction, but low cell density nonaggregated transit-amplifying cells still survived because Notch-1 is able to signal cell autonomously. In contrast, when medium Ca2+ is raised to >400 micromol/L, Notch-1 conformationally is no longer constitutively active but requires cell-cell contact for reciprocal binding of Jagged-1 ligands and Notch-1 receptors between adjacent transit-amplifying cells to activate their survival signaling. Such cell-cell contact is enhanced by the elevated Ca2+ inducing an E-cadherin conformation allowing homotypic interaction between transit-amplifying cells. Such Ca(2+)-dependent, E-cadherin-mediated interaction, however, results in cell aggregation, stratification, and inhibition of proliferation of transit-amplifying cells via contact inhibition-induced up-regulation of p27/kip1 protein. In addition, transit-amplifying cells not contacting other cells undergo squamous differentiation into cornified (i.e., 1% SDS insoluble) envelopes and death in the elevated Ca2+ medium. Stratification and contact inhibition induced by elevated Ca2+ are dependent on E-cadherin-mediated homotypic interaction between transit-amplifying cells as shown by their prevention in the presence of a cell-impermanent, E-cadherin neutralizing antibody. In contrast to growth inhibition of normal transit-amplifying cells, supplementation of low-Ca(2+)-SFD medium with 10% FCS and raising the Ca2+ to >600 micromol/L stimulates the growth of all prostate cancer cell lines tested. Additional results document that, at physiologic levels of Ca2+ (i.e., >600 micromol/L), prostatic cancer cells are not contact inhibited by E-cadherin interactions and Notch-1 signaling is no longer required for survival but instead becomes one of multiple signaling pathways for proliferation of prostatic cancer cells. These characteristic changes are consistent with prostate cancer cells' ability to metastasize to bone, a site of high-Ca2+ levels.
authors.affiliation: Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins and Departments of Urology and Pathology, Johns Hopkins School of Medicine, Baltimore, MD 21231-1000, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Dalrymple
authors.forename: Lizamma
authors.initials: L
authors.lastname: Antony
authors.forename: Yi
authors.initials: Y
authors.lastname: Xu
authors.forename: Aarti R
authors.initials: AR
authors.lastname: Uzgare
authors.forename: Julia T
authors.initials: JT
authors.lastname: Arnold
authors.forename: Jurga
authors.initials: J
authors.lastname: Savaugeot
authors.forename: Lori J
authors.initials: LJ
authors.lastname: Sokoll
authors.forename: Angelo M
authors.initials: AM
authors.lastname: De Marzo
authors.forename: John T
authors.initials: JT
authors.lastname: Isaacs
mesh_terms: D015820:Cadherins / Q000502:physiology*; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D048708:Cell Growth Processes / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D011467:Prostate / Q000166:cytology / Q000378:metabolism; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D051881:Receptor, Notch1 / Q000502:physiology*; D015398:Signal Transduction; D019098:Telomerase / Q000378:metabolism
chemical_list: D015820:Cadherins; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins; C497761:NOTCH1 protein, human; D051881:Receptor, Notch1; D019098:Telomerase; D002118:Calcium