The Targeted Disruption of Both Alleles of RARβ2 in F9 Cells Results in the Loss of Retinoic Acid-associated Growth Arrest

Abstract

F9 teratocarcinoma cell lines, carrying one or two disrupted alleles of the RARbeta(2) gene, were generated by homologous recombination to study the role of RARbeta(2) in mediating the effects of retinoids on cell growth and differentiation. Retinoic acid (RA) does not induce growth arrest of the RARbeta(2)-/- cells, whereas the F9 WT and RARbeta(2)+/- heterozygote lines undergo RA-induced growth arrest. The RARbeta(2)+/- lines also exhibit a faster cell cycle transit time in the absence of RA. The RARbeta(2)-/- stem cells exhibit an altered morphology when compared with the F9 WT parent line, and after RA treatment, the RARbeta(2)-/- cells do not exhibit a fully differentiated cell morphology. As compared with F9 WT cells, the RARbeta-/- cells exhibited a markedly lower induction of several early RA-responsive genes and no induction of laminin B1, a late response gene. The induction of RA metabolism in the F9 RARbeta(2)-/- cells following differentiation was not impaired. The research presented here, and prior research suggest that RARbeta is required for RA-induced growth arrest in a variety of cell types and that RARbeta also functions in mediating late responses to RA. These findings are significant in view of the reduced expression of RARbeta transcripts in a number of different types of human carcinomas.