Prolactin-Stimulated X-linked inhibitor of apoptosis protein expression during S phase cell cycle progression in rat...

The rat Nb2 lymphoma is useful for studying prolactin (PRL) receptor signaling to mitogenesis and apoptosis suppression. Previous results showed that PRL rapidly induced expression of several apoptosis suppressor genes during the G1 phase of cell cycle in this model. The X-linked inhibitor of apoptosis protein (XIAP) gene product acts to suppress apoptosis by direct inhibition of caspases. The present study was conducted to determine whether PRL alters XIAP expression in lactogen-dependent Nb2-11 or -independent Nb2-SFJCD1 cells. Stationary Nb2-11 cultures expressed detectable levels of an 8.9-kb XIAP transcript. PRL (20 ng/mL) stimulated its expression, reaching maximal levels within 12 h. Expression of XIAP was also evaluated in Nb2-SFJCD1 cells subsequent to treatment with differentiating agents (sodium butyrate [2 mM, 72 h], all trans-retinoic acid [10 microM, 72 h], or 1,25-dihydroxycholecalciferol [100 nM, 24 h]). PRL significantly increased XIAP expression in cells previously treated with these compounds. Further analysis revealed that PRL stimulated XIAP expression during S phase of the cell cycle. To determine whether XIAP suppressed apoptosis, its cDNA was stably transfected into Nb2-11 cells. Compared to controls, cells overexpressing XIAP exhibited substantially less DNA fragmentation when apoptosis was induced by PRL deprivation or glucocorticoids. We conclude that PRL-stimulated XIAP expression likely serves to suppress apoptosis as cells progress through the later phases of the cell cycle.