We recently reported that in ELT3 uterine leiomyoma cells, but not in normal myometrial cells, endothelin (ET)-1 exerts a survival effect insensitive to MAPK3/1(ERK1/2) inhibition. In the present work, we investigated the potential role of MAPK14 (p38) in this ET-1-mediated effect. We demonstrated that, in ELT3, but not in normal myometrial cells, ET-1 activated MAPK14. Data based on pharmacological and siRNA approaches indicate that ETA and ETB receptors contributed to the activation of MAPK14 by ET-1 through a mechanism involving Gi protein, but not PI3-kinase. The inhibition of MAPK3/1 by U0126 did not affect the activation of MAPK14 by ET-1. Conversely, the inhibition of MAPK14 by SB203580 and the down-regulation of MAP2K3/MAP2K6 (kinases upstream of MAPK14) by specific siRNA did not alter the activation of MAPK3/1. These data indicate that MAPK14 was activated by ET-1 independently from MAPK3/1. Furthermore, ET-1 increased protein expression of prostaglandin synthase 2 (PTGS2 or COX2), prostaglandin E2 (PGE2) production, and subsequent ELT3 cell survival. The inhibition of PTGS2 induction and subsequent survival induced by ET-1 required the coinhibition of MAPK14 and MAPK3/1. Our findings provide evidence that ET-1 activated MAPK14 only in ELT3 cells, but not in normal myometrial cells. This MAPK14 activation was required, in addition to MAPK3/1 in ET-1-mediated survival through the COX2/prostaglandin axis, and may explain the absence of ET-1 antiapoptotic effect in normal myometrial cells. Our data reinforce the role of ET-1 and associated signaling pathways in leiomyoma pathology.