PKCα-MAPK/ERK-phospholipase A2 signaling is required for human melanoma-enhanced brain endothelial cell proliferation and motility
PKCα-MAPK/ERK-phospholipase A2 signaling is required for human melanoma-enhanced brain endothelial cell proliferation and motility
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DOI：
10.1016/j.mvr.2009.09.001
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发表时间：
2009-12-01
影响因子：
3.1
通讯作者：
Alberghina, Mario
中科院分区：
文献类型：
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作者：
Anfuso, Carmelina Daniela;Giurdanella, Giovanni;Alberghina, Mario
研究人员对涉及肿瘤细胞相互作用和粘附的人黑色素瘤细胞（HMC）系和脑内皮细胞（EC）之间很大程度上未定义的信号转导机制和串扰进行了研究。在永生化大鼠脑 GP8.3 EC 培养物中，由 SK-MEL28 和 OCM-1 黑色素瘤细胞制备的条件培养基 (CM) 显着增强花生四烯酸释放、胞质磷脂酶 A(2) (cPLA(2)) 和 Ca+ 独立磷脂酶 A(2) (iPLA(2)) 比活性以及细胞生长 24 小时。诸如渥曼青霉素和 LY294002（相对于 PI3 激酶活性）、AACOCF(3)（相对于 cPLA(2) 和 iPLA(2)）、PD98059（相对于 ERK1/2 活性）和 NS-398（相对于环氧合酶-2 活性、COX-2）等抑制剂均能够阻断使用划痕伤口愈合测定确定的细胞增殖和运动性在黑色素瘤 CM 刺激的 EC 单层中。这些培养基还支持来自大鼠脑 (BBEC) 的原代 EC 的细胞增殖增强。将抗cPLA(2)抗体电穿孔至EC中可显着抑制响应CM的EC增殖。两种 CM 共孵育 24 小时后，cPLA(2)、PKC α、ERK1/2 的磷酸化、cPLA(2) 和 iPLA(2) 的蛋白和 mRNA 表达以及 COX-2 蛋白表达均显着刺激，并被特异性抑制剂减弱。通过共聚焦显微镜，可以清楚地观察到在 CM 刺激的培养物中生长的 EC 的核周和膜区域中 cPLA(2)、ERK1/2、PKC α 和 COX-2 的激活。因此，在 EC 培养物中，MEK-PKC α-ERK1/2 和 PI3-K/Akt 存活途径在与来自两种黑色素瘤细胞系的 CM 相互作用期间被激活，为理解 EC 代谢和信号转导提供了新的见解。这些途径代表了抑制或增强肿瘤血管生成的潜在治疗靶点。 (C) 2009 Elsevier Inc. 保留所有权利。
The largely undefined signal transduction mechanisms and cross-talk between human melanoma cell (HMC) lines and brain endothelial cells (ECs) involved in tumor cell interaction and adhesion were investigated. In immortalized rat brain GP8.3 EC cultures, conditioned media (CM) prepared from SK-MEL28 and OCM-1 melanoma cells significantly enhanced arachidonic acid release, cytosolic phospholipase A(2) (cPLA(2)) and Ca+-independent phospholipase A(2) (iPLA(2)) specific activities, and cell growth by 24 h. Inhibitors such as wortmannin and LY294002 (vs. PI3 kinase activity), AACOCF(3), (vs. cPLA(2) and iPLA(2)), PD98059 (vs. ERK1/2 activity) and NS-398 (vs. cyclooxygenase-2 activity, COX-2) were all able to block cell proliferation and motility determined using a scratch wound healing assay in melanoma CMs-stimulated EC monolayers. These media also support the enhanced cell proliferation of primary ECs derived from rat brain (BBEC). Electroporation of anti-cPLA(2) antibody into ECs markedly inhibited the EC proliferation in response to CMs. With both CMs, phosphorylation of cPLA(2), PKC alpha, ERK1/2, protein and mRNA expression of cPLA(2) and iPLA(2), and COX-2 protein expression were significantly stimulated after 24 h coincubation, and attenuated by specific inhibitors. By confocal microscopy, activation of cPLA(2), ERK1/2, PKC alpha and COX-2 in perinuclear and membrane regions of ECs grown in CM-stimulated cultures were clearly observed. Thus MEK-PKC alpha-ERK1/2 and PI3-K/Akt survival pathways are activated in EC cultures during the interaction with CM from both melanoma cell lines, providing new insight in understanding EC metabolism and signaling. These pathways represent potential therapeutic targets to inhibit or enhance tumor angiogenesis. (C) 2009 Elsevier Inc. All rights reserved.