Institut de Biochimie et Biophysique Moléculaire et Cellulaire, CNRS/UMR 8619, Université Paris-Sud 11, Orsay, France.
Biol Reprod. 2011 Mar;84(3):495-504. doi: 10.1095/biolreprod.110.089011. Epub 2010 Nov 17.
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.
我们最近报道称,在 ELT3 子宫平滑肌瘤细胞中,内皮素(ET)-1 发挥生存作用,而不受 MAPK3/1(ERK1/2)抑制的影响,但在正常子宫平滑肌细胞中不受影响。在本工作中,我们研究了 MAPK14(p38)在这种 ET-1 介导的作用中的潜在作用。我们证明,在 ELT3 中,但在正常子宫平滑肌细胞中,ET-1 激活了 MAPK14。基于药理学和 siRNA 方法的数据表明,ETA 和 ETB 受体通过涉及 Gi 蛋白但不涉及 PI3-激酶的机制,为 ET-1 激活 MAPK14 做出贡献。U0126 抑制 MAPK3/1 不影响 ET-1 对 MAPK14 的激活。相反,SB203580 抑制 MAPK14 和特异性 siRNA 下调 MAP2K3/MAP2K6(MAPK14 的上游激酶)不改变 MAPK3/1 的激活。这些数据表明,MAPK14 被 ET-1 激活,而不依赖于 MAPK3/1。此外,ET-1 增加了前列腺素合酶 2(PTGS2 或 COX2)、前列腺素 E2(PGE2)的产生以及随后的 ELT3 细胞存活。PTGS2 诱导和随后的 ET-1 诱导的存活的抑制需要 MAPK14 和 MAPK3/1 的共同抑制。我们的研究结果提供了证据表明,ET-1 仅在 ELT3 细胞中而不在正常子宫平滑肌细胞中激活 MAPK14。这种 MAPK14 激活除了 MAPK3/1 在 ET-1 介导的存活中通过 COX2/前列腺素轴起作用外,还需要 MAPK14 激活,这可能解释了 ET-1 在正常子宫平滑肌细胞中没有抗凋亡作用。我们的数据强化了 ET-1 和相关信号通路在平滑肌瘤病理中的作用。