Effects of 1,25-dihydroxyvitamin D3 on transformed C3H/10T1/2 fibroblasts grown as multicellular spheroids

We have previously shown that 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3) is able to modulate proliferation of C3H/10T1/2 fibroblasts grown in monolayer cell culture. The aim of this study was to investigate the effects of the hormone in a 3-dimensional culture system, which is considered to better reflect the cellular environment that is found in vivo. Two transformed cell lines of C3H/10T1/2 fibroblasts (C116 and TPA 482) were grown as multicellular spheroids, whereas the nontumorigenic C3H/10T1/2 C18 cells failed to form spheroids. The hormone had no effect on spheroids made of TPA 482 cells but had a biphasic effect on C116 spheroid growth with stimulation at low and inhibition at higher concentrations. In C116 spheroids, 10(-8) M 1,25-(OH)2D3 induced complete arrest of spheroid growth. Light and electron microscopic studies and immunocytochemistry showed that 10(-8) M 1,25-(OH)2D3 induced morphological changes in C116 spheroids, with a marked reduction in expression of fibronectin, laminin and the gap junction protein connexin 43. These effects were followed by a loosening of the spheroid structure, detachment of viable cells, and finally disaggregation of the spheroids. The results of this study, together with the data previously obtained in monolayer cell cultures, indicate that the effects of 1,25-(OH)2D3 may be conditioned by the cellular environment. Therefore, the spheroid model is an interesting tool for investigation of the antineoplastic effect of the hormone.