Journal of Lipid Research, Vol. 20, 357-362, March 1979
Copyright © 1979 by Lipid Research, Inc.

Retinoid metabolism in spontaneously transformed mouse fibroblasts (Balb/c 3T12-3 cells): enzymatic conversion of retinol to anhydroretinol

Pangala V. Bhat , Luigi M. De Luca , Sergio Adamo , Irene Akalovsky , Carol S. Silverman-Jones , and Gary L. Peck

Dermatology Branch and Differentiation Control Section, National Cancer Institute, Bethesda, MD 20014

Spontaneously transformed mouse fibroblasts (Balb/c 3T12-3 cells) displayed an increased adhesion when cultured in the presence of 10^-6 M all-trans retinol and acquired morphological characteristics of the normal pheno-type. Thus it was of interest to investigate the metabolism of [15-¹⁴C]retinol in this system. Within 24 hours of culture, approximately 4.25% of the [¹⁴C]retinol was taken up by the cells. The hydrocarbon [¹⁴C]anhydroretinol was a major metabolic product and was identified by gas-liquid chromatography and by its typical ultraviolet absorption spectrum with maxima at 386, 364, and 346 nm. At 24 and 40 hours anhydroretinol represented 27% and 55%, respectively, of the total nonpolar metabolites or approximately 16% and 30% of the total radioactive products. Formalin-fixed fibroblasts or cultured intestinal mucosal cells did not convert retinol into anhydroretinol. A more polar product with a UV absorption maximum at 310 nm was also found. The time course of the synthesis of this product by 3T12 cells suggested a precursor-product relationship with anhydroretinol. A microsomal preparation from 3T12 cells was also active in synthesizing [¹⁴C]anhydroretinol and [¹⁴C]metabolite-310 from [¹⁴C]retinol. Moreover incubation of metabolite-310 with the 3T12 microsomes yielded anhydroretinol (40% conversion in 30 minutes), suggesting that metabolite-310 is an intermediate in the synthesis of anhydroretinol by these cells. Anhydroretinol appears to be an end product of the metabolism of retinol in 3T12-3 cells, as suggested by the finding that over 90% of [¹⁴C]anhydroretinol incubated for 30 hours with 3T12-3 cells was recovered unaltered, without the formation of detectable retroretinol, retinol, or retinoic acid.—Bhat, P. V., L. M. De Luca, S. Adamo, I. Akalovsky, C. S. Silverman-Jones, and G. L. Peck. Retinoid metabolism in spontaneously transformed mouse fibroblasts (Balb/c 3T12-3 cells): enzymatic conversion of retinol to anhydroretinol.

Supplementary key words microsomal system • 3T3 mouse fibroblasts • primary dermal fibroblasts • intestinal mucosal cells • vitamin A

Submitted on May 31, 1978
Accepted on October 20, 1978

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