Abstract：
Platelets can support cancer progression via the release of microparticles and microvesicles that enhance the migratory behaviour of recipient cancer cells. We recently showed that platelet-derived extracellular vesicles (PEVs) stimulate migration and invasiveness in highly metastatic MDA-MB-231 cells by stimulating the phosphorylation of p38 MAPK and the myosin light chain 2 (MLC2). Herein, we assessed whether the pro-migratory effect of PEVs involves the remodelling of the Ca2+ handling machinery, which drives MDA-MB-231 cell motility.
PEVs were isolated from human blood platelets, and Fura-2/AM Ca2+ imaging, RT-qPCR, and immunoblotting were exploited to assess their effect on intracellular Ca2+ dynamics and Ca2+-dependent migratory processes in MDA-MB-231 cells.
Pretreating MDA-MB-231 cells with PEVs for 24 h caused an increase in Ca2+ release from the endoplasmic reticulum (ER) due to the up-regulation of SERCA2B and InsP3R1/InsP3R2 mRNAs and proteins. The consequent enhancement of ER Ca2+ depletion led to a significant increase in store-operated Ca2+ entry. The larger Ca2+ mobilization from the ER was required to potentiate serum-induced migration by recruiting p38 MAPK and MLC2.
PEVs stimulate migration in the highly metastatic MDA-MB-231 breast cancer cell line by inducing a partial remodelling of the Ca2+ handling machinery.
PEVs were isolated from human blood platelets, and Fura-2/AM Ca2+ imaging, RT-qPCR, and immunoblotting were exploited to assess their effect on intracellular Ca2+ dynamics and Ca2+-dependent migratory processes in MDA-MB-231 cells.
Pretreating MDA-MB-231 cells with PEVs for 24 h caused an increase in Ca2+ release from the endoplasmic reticulum (ER) due to the up-regulation of SERCA2B and InsP3R1/InsP3R2 mRNAs and proteins. The consequent enhancement of ER Ca2+ depletion led to a significant increase in store-operated Ca2+ entry. The larger Ca2+ mobilization from the ER was required to potentiate serum-induced migration by recruiting p38 MAPK and MLC2.
PEVs stimulate migration in the highly metastatic MDA-MB-231 breast cancer cell line by inducing a partial remodelling of the Ca2+ handling machinery.
摘要：
血小板可以通过释放增强受体癌细胞迁移行为的微粒和微泡来支持癌症进展。我们最近表明，血小板衍生的细胞外囊泡（PEV）通过刺激p38MAPK和肌球蛋白轻链2（MLC2）的磷酸化来刺激高转移性MDA-MB-231细胞的迁移和侵袭性。在这里，我们评估了PEV的促迁移作用是否涉及Ca2处理机械的重塑，驱动MDA-MB-231细胞运动。
PEV是从人的血小板中分离出来的，和Fura-2/AMCa2+成像，RT-qPCR,并利用免疫印迹来评估其对MDA-MB-231细胞中细胞内Ca2动力学和Ca2依赖性迁移过程的影响。
用PEV预处理MDA-MB-231细胞24小时，由于SERCA2B和InsP3R1/InsP3R2mRNA和蛋白质的上调，导致内质网（ER）释放的Ca2增加。ERCa2消耗的增加导致存储操作的Ca2进入显着增加。通过募集p38MAPK和MLC2，需要从ER动员更大的Ca2才能增强血清诱导的迁移。
PEV通过诱导Ca2处理机制的部分重塑来刺激高度转移性MDA-MB-231乳腺癌细胞系中的迁移。
PEV是从人的血小板中分离出来的，和Fura-2/AMCa2+成像，RT-qPCR,并利用免疫印迹来评估其对MDA-MB-231细胞中细胞内Ca2动力学和Ca2依赖性迁移过程的影响。
用PEV预处理MDA-MB-231细胞24小时，由于SERCA2B和InsP3R1/InsP3R2mRNA和蛋白质的上调，导致内质网（ER）释放的Ca2增加。ERCa2消耗的增加导致存储操作的Ca2进入显着增加。通过募集p38MAPK和MLC2，需要从ER动员更大的Ca2才能增强血清诱导的迁移。
PEV通过诱导Ca2处理机制的部分重塑来刺激高度转移性MDA-MB-231乳腺癌细胞系中的迁移。
