The antagonistic role of activated oncogenes and apoptosis : studies of n-myc and P53 in lymphoid tumorigenesis

Abstract: Inactivation of the p53 tumor suppre.ssor gene and activation of proto-oncogenes have been implicated in the initiation and/or progression of anumber of neoplasms. In this thesis, the involvement of N-myc gene activationhas been studied in mouse plasmacytomagenesis and the role of wild-type (wt)p53 and oncogene activation has been evaluated with particular emphasis onapoptosis .Nearly all mouse plamacytomas (MPC) carry Ig/c-myc translocations. We showedthat MPCs could be induced in Emy-N-myc transgenic mice. All the tumorsexpressed the N-myc transgene but not c-myc. None of them carried the MPC-associated Ig/c-myc translocations. In the conventional MPC-induction system,an exceptional MPC with a novel t(6;12) translocation was identified thatjuxtaposed N-myc to the Igk locus. A functional equivalence between c-myc andN-myc in plasmacytomagenesis was suggested.We have found that a v-myc retrovirus-induced T cell Iymphoma line (J3D) haslost one of its p53 alleles, whereas the other has become inactivated due to aninsertion of a Moloney murine leukemia provirus in intron 4 with an oppositetranscriptional orientation. No p53 protein could be detected. Reconstitution ofthese cells with wt p53 using a temperature-sensitive (ts) murine Val135 mutantconstruct demonstrated that wt p53 can trigger both G1 cell cycle arrest andapoptosis. A kinetic analysis showed that wt p53-induced G1 arrest precededapoptosis. We have hypothesized that apoptosis is triggered by the contradictorysignals of the growth-promoting activated myc and the growth-inhibiting wtp53.Introduction of a bcl-2 expression vector into the J3D ts p53 cells inhibited wtp53-induced apoptosis. However, bc1-2 had no effect on wt pS3-induced Gl arrestand did not interfere with wt p53-mediated transactivation Of p21WAFl and bax.We suggest that inhibition of p53-induced apoptosis by bcl-2 could be due toneutralization of the apoptosis-promoting effect of bax. These findings areconsistent with the possibility that p21WAFl induction is responsible for pS3-mediated Gl arrest and that p53-mediated induction of bax accelerates apoptosis.An expression vector carrying the human papillomavirus (HPV) 16 E7 gene wasintroduced into the ts p53 J3D cells. A partial relief from p53-induced Gl arrestby overexpression of E7 affected neither the kinetics nor the frequency of p53-triggered apoptosis. Expression of E7 protein did not interfere with pS3-mediatedtransactivation of p21WAFl and bax. This indicates that pS3-induced apoptosis isindependent of p53-induced Gl arrest.Key Words: MPC, EmyN-myc transgene, J3D T Iymphoma cells, p53, v-m y cretrovirus, bcl-2, Gl cell cycle arrest, apoptosis, HPV 16 E7ISBN: 91-628-1878-3