Neoplastic phenotype of gap-junctional intercellular communication-deficient WB rat liver epithelial cells and its reversal by forced expression of connexin 32.
Gap-junctional intercellular communication (GJIC) is involved in cellular growth control and is often reduced in neoplastic cells. In this study, four GJIC-deficient rat liver epithelial cell lines (WB-aB1, WB-bA2, WB-cD6, and WB-dA2) were examined for altered growth and tumorigenicity in comparison with their GJIC-competent parental cell line, WB-F344. WB-aB1 cells were also forced to express connexin 32 (Cx32) by transduction with a Cx32 cDNA retroviral expression vector to help determine whether the restoration of GJIC could reverse their neoplastic phenotype. WB-aB1 and WB-bA2 cells had faster population doubling times (PDTs) and higher saturation densities (SDs) than did WB-F344 cells. In contrast, the growth of WB-cD6 and WB-dA2 cells was not significantly different from that of WB-F344 cells. WB-aB1 and WB-bA2 cells formed tumors in male F344 rats, but WB-cD6 and WB-dA2 cells did not. After transduction of WB-aB1 cells with Cx32, four stable clones (WB-a/32-3, -8, -9, and -10) were isolated that had GJIC levels of 5.2%, 44.5%, 69.8%, and 90.5%, respectively. The growth of poorly coupled clones 3 and 8 was similar to that of parental WB-aB1 cells, but the growth of well-coupled clones 9 and 10 was similar to that of WB-F344 cells. The tumorigenicity of WB-a/32-9 and WB-a/32-10 cells was also significantly lower than that of WB-aB1 cells. Our results suggest that reduced GJIC contributes to neoplastic transformation of WB cells, that additional changes are necessary, and that restoration of GJIC by forced Cx32 protein expression can suppress the neoplastic phenotype of these cells.
Rae
R S
RS
Department of Pathology, Medical College of Ohio, Toledo 43699, USA.
Mehta
P P
PP
Chang
C C
CC
Trosko
J E
JE
Ruch
R J
RJ
D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002471:Cell Transformation, Neoplastic; D017630:Connexins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D017629:Gap Junctions / Q000502:physiology*; D008099:Liver / Q000166:cytology* / Q000378:metabolism* / Q000502:physiology; D008297:Male; D009154:Mutation; D010641:Phenotype; D051381:Rats; D011916:Rats, Inbred F344; D014161:Transduction, Genetic; D014162:Transfection; D000097002:Gap Junction beta-1 Protein
D017630:Connexins; D000097002:Gap Junction beta-1 Protein