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Induction of AP-1 activity associated with c-Jun and JunB is required for mitogenesis induced by insulin and vanadate in SV40-transformed 3T3T cells

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Abstract

Insulin and vanadate function as complete mitogens for SV40-transformed murine 3T3T (CSV3-1) cells but not for nontransformed 3T3T cells. Mitogenesis induced by insulin and vanadate in CSV3-1 cells is associated with the induction of the expression of protooncogenes c-Jun and junB, two major AP-1 transcription factor components. We now report that both insulin and vanadate induce a significant increase in AP-1 DNA binding activity in CSV3-1 cells but not in 3T3T cells. Gel supershift assays and Western blot analysis using specific antibodies demonstrate that the increased AP-1 binding activity induced by insulin and vanadate in CSV3-1 cells is primarily contributed by an increase in the expression of c-Jun and JunB protein levels. Furthermore, treatment of CSV3-1 cells with antisense oligodeoxyribonucleotides to c-Jun or to junB blocks insulin- and vanadate-induced mitogenesis whereas antisense junD oligomers have no inhibitory effects. These results therefore demonstrate that the induction of AP-1 binding activity associated with c-Jun and JunB is required for insulin- and vanadate-induced rnitogenesis in SV40-transformed murine 3T3T cells. Additional data presented in this paper show that JunD/AP-1 binding activity, which is thought to play a negative role in regulating cell proliferation, is also slightly induced following insulin and vanadate stimulation in CSV3-1 cells. Nevertheless, the ratio of proliferation promoting c-Jun/AP-1 and JunB/AP-1 binding activities to proliferation inhibiting JunD/AP-1 binding activity is significantly increased following insulin and vanadate stimulation. These results therefore support the concept that modulation of the balance of positive Jun/AP-1 and negative Jun/AP-1 activities is important in regulating cell proliferation.

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Authors and Affiliations

  1. Department of Pathology, The University of Tennessee College of Medicine, 899 Madison Avenue, Memphis, Tennessee

    Hanlin Wang, Zirong Xie & Robert E. Scott

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  1. Hanlin Wang
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  2. Zirong Xie
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  3. Robert E. Scott
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Wang, H., Xie, Z. & Scott, R.E. Induction of AP-1 activity associated with c-Jun and JunB is required for mitogenesis induced by insulin and vanadate in SV40-transformed 3T3T cells. Mol Cell Biochem 168, 21–30 (1997). https://doi.org/10.1023/A:1006889623326

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