Increased fucosylation of high‐molecular‐weight glycoproteins accompanies retinoic‐acid‐induced differentiation of f‐9 embryonal carcinoma cells

Abstract

Retinoic acid (RA) treatment of F-9 embryonal carcinoma cells resulted in cell flattening and increased production of laminin B1 chain, both indicating differentiation to endoderm-like cells. In addition, RA caused a time- and dose-dependent decrease in growth rate in monolayer culture and a dose-dependent decrease in the ability of the cells to form colonies in soft agarose. Differentiation was accompanied by an increase in the fucosylation of specific high-molecular-weight cellular and cell-surface glycoproteins. The fucosylation of glycoproteins of Mr 175,000 (gp175), 250,000 (gp250), and 400,000 (gp400) increased as early as 24 hr after the addition of 5 x 10(-6) M RA to the culture medium. These changes preceded both growth inhibition and the induction of laminin B1 expression, which were detected 48 to 72 hr after addition of RA. The increased fucosylation of these glycoproteins showed a distinct dose-response relationship. Both gp175 and gp250 showed the greatest increase in fucosylation at 10(-5) M, which was also the dose at which RA induced laminin maximally, while the fucosylation of gp400 was greatest at 10(-8) M RA and declined at higher concentrations. The overall synthesis of large fucosylated glycopeptides decreased in RA-treated cells, in spite of the increases in the fucosylation of specific cellular glycoproteins. RA-induced differentiation of F-9 cells was also accompanied by a time- and dose-dependent increase in fucosyltransferase activity. Although the functions of these glycoproteins are not currently known, the early increase in their fucosylation can be considered as a marker of differentiation in this system.

Effect of retinoic acid on cell proliferation and differentiation as well as on lipid synthesis, lipoprotein secretion, and apolipoprotein biogenesis

“…Similarly, a number of in vivo and in vitro studies using various experimental models have revealed that RA plays a key role in embryonic development, the prevention and treatment of tumors, as well as cell proliferation and differentiation (10,12). In particular, our data are in line with many reports suggesting that RA is more active in inhibiting proliferation and inducing differentiation in rela- tively undifferentiated systems, i.e., murine F9 teratocarcinoma cells, promiolocytic HL-60 cells, L-6 melanoma cells, and neuroblastoma and embryonal stem cells P19 (2,20,49). In contrast, in other cell lines, such as keratinocytes (66), embryonal lung cells (63), and chick embryo hepatocytes (70), proliferation is stimulated.…”

α-L-Fucose: A Potentially Critical Molecule in Pathologic Processes Including Neoplasia

“…Retinoic acid-induced differentiation of F9 embryonal carcinoma cells has been accompanied by increased fucosyltransferase activity, as well as by increased fucosylation of certain specific high molecular weight cell-surface glycoproteins. 168 Similar effects were seen with retinoic acid treatment of murine melanoma cells. 169 The meaning of these discrepant findings is unclear.…”

Transcriptional Regulation of α1,3-Galactosyltransferase in Embryonal Carcinoma Cells by Retinoic Acid

“…Treatment of F9 cells for 5 days with RA and dibutyryl cAMP results in an approximately 80% decline in the activity of an ␣1,3FT (28). However, using a different assay for fucosyltransferase activity another group observed a slight increase in the activity of fucosyltransferase upon RA treatment of F9 cells (88). In the case of ␤1,4GT, it was shown that ␤1,4GT activity in F9 cells declines after 3 days of differentiation and then begins to rise around 5-6 days of differentiation, which correlates with increased levels of transcript observed after 5-6 days of treatment with RA (53).…”