title: Connexin expression and cell coupling fail to reverse the v-src transformed growth characteristics of a Cx43-/- cell line.
abstract: Gap junctions, composed of connexins, have been shown to suppress transformation in a variety of malignancies and transformed cell types. In addition, transforming factors such as the src oncogene have been shown to directly phosphorylate some connexins (e.g., Cx43) and inhibit coupling. To investigate the role of gap junctions in cell transformsation by v-src, we utilized a clonal cell line derived from Cx43 knockout mice (KoA) that was immortalized, but not transformed. Transfection by v-src induced a marked transformed phenotype characterized by growth in low serum and anchorage-independent conditions. Subsequent transfections by Cx43, Cx32 or vector alone were then tested for their effects on growth. Activity of pp60v-src was confirmed in all transfectants as well as the ability of pp60v-src to phosphorylate Cx43 in several clones. Despite the documented effect of pp60v-src on Cx43 channel closure, modest coupling was still retained in many of the Cx43 and Cx32 transfectants. However, none of the four Cx43 transfected clones showed significant inhibitory effects on proliferation in either anchorage-independent or low serum growth conditions. Of the Cx32 clones, only one in five showed effects on growth in both assays, which was the same ratio observed for the control transfectants. Thus, based on the levels of expression achieved, which were comparable to endogenous levels in established cell lines, neither Cx43 nor Cx32 serve as effective suppressors of the transformed growth phenotype of this v-src expressing cell line.
authors.affiliation: Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.
authors.forename: A
authors.initials: A
authors.lastname: Chandrasekhar
authors.forename: M
authors.initials: M
authors.lastname: Merritt
authors.forename: S J
authors.initials: SJ
authors.lastname: Huh
authors.forename: B J
authors.initials: BJ
authors.lastname: Nicholson
authors.forename: S N
authors.initials: SN
authors.lastname: Zucker
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000502:physiology*; D002450:Cell Communication; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D018031:Connexin 43 / Q000172:deficiency / Q000235:genetics*; D017630:Connexins / Q000235:genetics; D003470:Culture Media; D017353:Gene Deletion; D016391:Genes, src / Q000235:genetics*; D005822:Genetic Vectors; D051379:Mice; D018345:Mice, Knockout; D014162:Transfection; D000097002:Gap Junction beta-1 Protein
chemical_list: D018031:Connexin 43; D017630:Connexins; D003470:Culture Media; D000097002:Gap Junction beta-1 Protein