Inhibition of intrinsic gap-junction intercellular communication and enhancement of tumorigenicity of the rat bladder carcinoma cell line BC31 by a dominant-negative connexin 43 mutant.
The tumor-suppressive property of the connexin gap-junction proteins was postulated from the fact that their function of cell coupling is impaired in most cancer cells. However, in conflict with this notion, certain cancer cells are able to communicate through gap junctions despite their malignancy. To explain this phenomenon, we studied by using a dominant-negative strategy the effect on tumorigenicity of loss of intrinsic gap-junction intercellular communication (GJIC) in the rat bladder carcinoma cell line BC31, which shows both expression of connexin 43 (Cx43) and intercellular communication. In cells transfected with a mutant Cx43 with seven residues deleted from the internal loop at positions 130-136 (Cx43delta), transport of the resulting connexin protein to the plasma membrane occurred normally, but the GJIC of the cells was effectively abolished at the level of permeability of established gap junctions. Dominant-negative inhibition of GJIC by Cx43delta accelerated growth of BC31 cells in nude mice. In contrast, when GJIC in BC31 cells was artificially enforced by transfection of wild-type Cx43, the cells lost the capacity to grow in vivo. Decreased phosphorylation of Cx43delta suggested close interaction of the internal loop of connexin with its commonly phosphorylated domains in the C-terminal tail and involvement of this interaction in gap-junction permeability. Therefore, we conclude that the intrinsic GJIC observed in cancer cells should be considered a tumor-suppressor factor and that its level may influence malignant growth capacity.
Krutovskikh
V A
VA
International Agency for Research on Cancer, Lyon, France.
Yamasaki
H
H
Tsuda
H
H
Asamoto
M
M
D000818:Animals; D002295:Carcinoma, Transitional Cell / Q000235:genetics* / Q000473:pathology; D002450:Cell Communication / Q000235:genetics*; D004396:Coloring Agents / Q000378:metabolism; D018031:Connexin 43 / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D017629:Gap Junctions / Q000502:physiology*; D005799:Genes, Dominant; D007546:Isoquinolines / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D016297:Mutagenesis, Site-Directed; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D009368:Neoplasm Transplantation; D011487:Protein Conformation; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology
D004396:Coloring Agents; D018031:Connexin 43; D007546:Isoquinolines; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; C017475:lucifer yellow