Pon Yuen L, Zhou Hong Y, Cheung Annie N Y, Ngan Hextan Y S, Wong Alice S T
School of Biological Sciences, University of Hong Kong, Hong Kong.
Cancer Res. 2008 Aug 15;68(16):6524-32. doi: 10.1158/0008-5472.CAN-07-6302.
p70 S6 kinase (p70(S6K)) is a downstream effector of phosphatidylinositol 3-kinase and is frequently activated in human ovarian cancer. Here we show that p70(S6K) functions in epithelial to mesenchymal transition (EMT) responsible for the acquisition of invasiveness during tumor progression. This tumorigenic activity is associated with the ability of p70(S6K) to repress E-cadherin through the up-regulation of Snail. p70(S6K) activation induced phenotypic changes consistent with EMT in ovarian cancer cells: The cells lost epithelial cell morphology, acquired fibroblast-like properties, and showed reduced intercellular adhesion. Western blot showed that p70(S6K) activation led to decreased expression of the epithelial marker E-cadherin and increased expression of mesenchymal markers N-cadherin and vimentin. Inhibition of p70(S6K) by a specific inhibitor or small interfering RNA reversed the shift of EMT markers. Importantly, p70(S6K) activation also stimulated the expression of Snail, a repressor of E-cadherin and an inducer of EMT, but not other family members such as Slug. This induction of Snail was regulated at multiple levels by increasing transcription, inhibiting protein degradation, and enhancing nuclear localization of Snail. RNA interference-mediated knockdown of Snail suppressed p70(S6K)-induced EMT, confirming that the effect was Snail specific. Furthermore, phospho (active)-p70(S6K) staining correlated with higher tumor grade. We also showed a significant positive correlation between p70(S6K) activation and Snail expression in ovarian cancer tissues. These results indicate that p70(S6K) may play a critical role in tumor progression in ovarian cancer through the induction of EMT. Targeting p70(S6K) may thus be a useful strategy to impede cancer cell invasion and metastasis.
p70核糖体蛋白S6激酶(p70(S6K))是磷脂酰肌醇3激酶的下游效应分子,在人类卵巢癌中经常被激活。在此我们表明,p70(S6K)在上皮-间质转化(EMT)过程中发挥作用,而EMT在肿瘤进展过程中负责侵袭性的获得。这种致瘤活性与p70(S6K)通过上调Snail来抑制E-钙黏蛋白的能力有关。p70(S6K)激活诱导了与卵巢癌细胞中EMT一致的表型变化:细胞失去上皮细胞形态,获得成纤维细胞样特性,并表现出细胞间黏附减少。蛋白质印迹法显示,p70(S6K)激活导致上皮标志物E-钙黏蛋白表达降低,间充质标志物N-钙黏蛋白和波形蛋白表达增加。用特异性抑制剂或小干扰RNA抑制p70(S6K)可逆转EMT标志物的转变。重要的是,p70(S6K)激活还刺激了Snail的表达,Snail是E-钙黏蛋白的抑制因子和EMT的诱导因子,但不刺激其他家族成员如Slug的表达。Snail的这种诱导在多个水平上受到调控,包括增加转录、抑制蛋白质降解以及增强Snail的核定位。RNA干扰介导的Snail敲低抑制了p70(S6K)诱导的EMT,证实该效应具有Snail特异性。此外,磷酸化(活性)-p70(S6K)染色与更高的肿瘤分级相关。我们还显示卵巢癌组织中p70(S6K)激活与Snail表达之间存在显著正相关。这些结果表明,p70(S6K)可能通过诱导EMT在卵巢癌的肿瘤进展中起关键作用。因此,靶向p70(S6K)可能是阻碍癌细胞侵袭和转移的一种有用策略。