title: Frog PNKT-4B cells express specific extracellular matrix-degrading enzymes and cytokines correlated with an invasive phenotype.
abstract: A temperature-dependent metastatic phenotype reported for a frog cell line, PNKT-4B, provided a means for studying potential mediators of cell-matrix interaction involved in metastatic invasion. Zymography revealed that these cells secreted enzyme species with properties and characteristics of mammalian metalloproteinases: collagenase, stromelysin, gelatinase A, and gelatinase B. These enzymes were produced by PNKT-4B cultures maintained at both invasive-permissive (28 degrees C), and invasion-restrictive (20 degrees C) temperatures. However, under the invasive-permissive culture condition cells produced more of the putative gelatinase B and A enzymes. In addition, an activated form of gelatinase A was produced only in invasion-permissive cultures. DNA synthesis bioassays (Mv1Lu cell line and mouse thymocytes) to detect growth promoting and/or inhibitory cytokines, revealed that PNKT-4B cultures kept at 28 degrees C released significantly higher levels of stimulatory (interleukin-1-like) and latent inhibitory (transforming growth factor-beta-like) substances into the medium compared to 20 degrees C cultures. Pre-absorption of media samples with heparin-sepharose indicated a second stimulatory cytokine as well. A corneal fibroblast bioassay that tests for mediators of collagenase synthesis, detected a stimulatory substance whose activity was greatly reduced in the presence of interleukin-1 receptor antagonist protein. Collagenase stimulatory activity present in 28 degrees C culture medium was significantly higher than equal samples from 20 degrees C cultures. These studies provide a molecular correlation between release of cytokines with properties of the metastatic phenotype seen in vivo. They further provide some of the first characterizations of frog MMPs and cytokines, which are likely to be involved in other tissue remodeling events.
authors.affiliation: Vision Research Laboratories, New England Medical Center, Boston Massachusetts 02111, USA.
authors.forename: K J
authors.initials: KJ
authors.lastname: Strissel
authors.forename: M J
authors.initials: MJ
authors.lastname: Tessier
authors.forename: N B
authors.initials: NB
authors.lastname: Shams
authors.forename: S
authors.initials: S
authors.lastname: Grabbe
authors.forename: J
authors.initials: J
authors.lastname: Gross
authors.forename: M E
authors.initials: ME
authors.lastname: Fini
mesh_terms: D000818:Animals; D001001:Anura; D002452:Cell Count; D017364:Collagenases / Q000096:biosynthesis / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D004247:DNA / Q000096:biosynthesis; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D008666:Metalloendopeptidases / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D009361:Neoplasm Invasiveness; D010641:Phenotype; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D004247:DNA; D017364:Collagenases; D008666:Metalloendopeptidases