Regulation of Par-4 by Oncogenic Ras

Oncogenic Ras causes down-regulation of the proapoptotic tumor suppressor gene Par-4. Replenishment of the basal levels of Par-4 results in inhibition of Ras-inducible cellular transformation. Moreover, overexpression of Par-4 (twofold to fourfold over basal levels) results in apoptosis of cells expressing oncogenic Ras. Par-4 does not, on its own, induce apoptosis in immortalized or nontransformed cells. This chapter describes the key methods used for analysis of Par-4 down-regulation by oncogenic Ras, which can be extended to study most genes whose down-regulation by oncogenic Ras is critical for oncogenic transformation and cell survival.