GROWTH-HORMONE INDUCES C-FOS AND C-JUN EXPRESSION IN CELLS WITH VARYING REQUIREMENTS FOR DIFFERENTIATION

GH is one of the major factors required for the differentiation of 3T3-F442A preadipocyte fibroblasts into adipocytes. An early event following the addition of GH to 3T3-F442A preadipocytes is induction of the expression of c-fos and c-jun. Although c-fos and c-jun expression has been observed in conjunction with growth factor-stimulated differentiation in several cell types, it is not clear whether protooncogene expression and differentiation are necessarily related. In this study the relationship between the induction of these protooncogenes and differentiation was evaluated by taking advantage of several cell lines that are related to 3T3-F442A cells but have varying GH requirements for differentiation. Adipose differentiation in the adipogenic cell lines 3T3-L1 and 3T3-GI-16 is known to be GH independent, requiring insulin or insulin-like growth factor-I. In both 3T3-L1 and 3T3-GI-16 preadipocytes, GH, nevertheless, induced the expression of mRNA for both protooncogenes. GH (2.2 nM) was more effective than insulin (1-mu-M) in inducing c-fos expression in these two adipogenic cell lines, suggesting that induction of the protooncogenes is not sufficient to induce adipogenesis. 3T3-C2 fibroblasts do not differentiate in response to any of the stimuli that convert 3T3-F442A fibroblasts to adipocytes. However, GH (2.2 nM) as well as calf serum induced the expression of c-fos and c-jun in 3T3-C2 cells. NIH-3T3 fibroblasts, which do not undergo differentiation, also showed induction of c-fos by GH. Thus, GH-induced expression of c-fos and c-jun occurs in nondifferentiating cells. Furthermore, in differentiated 3T3-F442A adipocytes, GH stimulated the expression of c-fos and c-jun as it does in the preadipocytes. Since GH elicits a variety of metabolic responses in 3T3-F442A adipocytes, the present findings raise the possibility that induction of c-fos and c-jun expression might be associated with multiple events in GH-stimulated 3T3-F442A adipocytes. The lack of requirement for GH in GH-independent and nondifferentiating cells compared to 3T3-F442A cells does not appear to reflect the lack of GH receptors, since expression of mRNA for the GH receptor was evident in all of the cell types tested and, thus, corresponds with the ability of GH to induce protooncogene expression. Although GH-induced c-fos expression was relatively invariant, since it was evident in all of the cell types studied, this response could clearly be regulated, since it was attenuated by prior exposure to GH or serum in 3T3-F442A preadipocytes. In this regard, this response is comparable to other short term responses to GH. Taken together, these studies indicate that while induction of c-fos and c-jun may well play a role in GH-induced differentiation in 3T3-F442A preadipocytes, this effect of GH is not exclusively associated with differentiation in 3T3-F442A and related cell types. Protooncogene expression may be an early event associated with other functions regulated by GH.