Transforming Growth Factor Beta Stimulates Mitogenically Mouse NIH3T3 Fibroblasts and Those Cells Transformed by the EJ-H-ras Oncogene

Abstract

TGF-β1 stimulates thymidine incorporation and the growth rate of mouse NIH3T3 fibroblasts and of those cells transformed by the EJ-H-ras oncogene (TR15 cells), in the presence and the absence of serum. Thymidine incorporation, in serum-deprived cells, is stimulated to a higher degree by 0.1-1 ng/ml of TGF-β in NIH3T3 than in TR15 cells, which have a 10-fold higher basal level of incorporation. In both cell types TGF-β1 is as active, or more active than other mitogens (TGF-α, PDGF-AB, bFGF) at the same concentration. The growth rate of NIH3T3 cells, in low serum or serum-free (S⁻) medium, is stimulated by only 10 picograms/ml of TGF-β1, and that of TR15 cells, in S⁻ medium, by only 1 picogram/m1. In contrast, TGF-β1 inhibits mitogenically unestablished mouse embryo fibroblasts and these fibroblasts immortalized spontaneously and able to grow in S⁻ medium. It also inhibits the anchorage-independent growth of TR15 cells. NIH3T3 and TR15 cells respond, similarly, to TGF-β activated by acification of their culture medium. The kinetics of thymidine incorporation and of activation of the c-myc proto-oncogene, observed already after 1 hr, in treated NIH3T3 and TR15 cells, suggests a direct mitogenic stimulation. The level of activated c-myc RNA is 2-fold higher at 2 hr, and subsequently decreases relatively less in the TR15 cells.

Key Words:

• transforming growth factor beta (TGF-β)
• NIH3T3 cells
• EJ-H-ras oncogene
• stimulation of growth