Role of Vimentin in the Inhibitory Effects of Low-Molecular-Weight Heparin on PC-3M Cell Adhesion to, and Migration through, Endothelium
Role of Vimentin in the Inhibitory Effects of Low-Molecular-Weight Heparin on PC-3M Cell Adhesion to, and Migration through, Endothelium
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波形蛋白在低分子量肝素对 PC-3M 细胞粘附和迁移通过内皮细胞的抑制作用中的作用
DOI：
10.1124/jpet.111.182055
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发表时间：
2011-10-01
影响因子：
3.5
通讯作者：
Li, Xuejun
中科院分区：
文献类型：
--
作者：
Pan, Yan;Lei, Tianluo;Li, Xuejun
低分子量肝素（LMWH）已被用于静脉血栓栓塞并发症的癌症患者，导致更高的生存率和对实验性转移的抑制作用。在本研究中，人脐静脉内皮细胞（HUVECs）与LMWH处理24小时。我们发现，所得的HUVECs可以显着抑制高转移性人前列腺癌细胞系（PC-3 M）在其粘附到内皮细胞和跨内皮细胞迁移，根据扫描电子显微镜。我们还测定了PC-3 M细胞与HUVECs的粘附通过与LMWH孵育大大降低后内皮细胞间Ca 2+浓度的升高水平。使用蛋白质组学，我们调查了低分子量肝素治疗后HUVEC的整体蛋白质变化，并确定了四种下调的蛋白质，它们可能是细胞骨架波形蛋白中间丝，软骨衍生的C型凝集素和丝氨酸/苏氨酸蛋白磷酸酶1 β（PP-1B）的亚型。LMWH可影响PC-3 M细胞与HUVECs结合后波形蛋白的形态、α（v）整合素和PP-1B的表达水平。波形蛋白有助于PC-3 M细胞的粘附，这通过短干扰RNA实验证实。此外，纯化的波形蛋白与LMWH直接结合的表面等离子体共振方法检测。然而，当我们使用荧光标记的肝素24小时，以确定这种结合是否发生在细胞内，肝素主要分布在内皮细胞周围。总之，这些发现表明，单孵育的LMWH与HUVECs可以抑制PC-3 M细胞粘附，并通过，内皮细胞迁移。LMWH对波形蛋白的调节在抗转移作用中起作用。
Low-molecular-weight heparin (LMWH) has been used in cancer patients with venous thromboembolic complications, resulting in a higher survival rate and an inhibitory action on experimental metastasis. In the present study, human umbilical vein endothelial cells (HUVECs) were treated with LMWH for 24 h. We found that the resulting HUVECs could significantly inhibit the highly metastatic human prostate cancer cell line (PC-3M) in terms of its adhesion to the endothelium and migration across the endothelium, according to scanning electron microscopy. We also determined the elevated levels of endothelial intercellular Ca2+ concentration after the adhesion of PC-3M cells to HUVECs was greatly reduced by incubation with LMWH. Using proteomics, we surveyed the global protein changes in HUVECs after LMWH treatment and identified four down-regulated proteins that were possible isoforms of cytoskeletal vimentin intermediate filaments, cartilage-derived C-type lectin, and serine/threonine protein phosphatase 1 beta (PP-1B). LMWH affected the morphology of vimentin and the expression levels of alpha(v) integrin and PP-1B in HUVECs bound to PC-3M cells. Vimentin assists in the adhesion of PC-3M cells, which was confirmed by short interfering RNA experiments. Furthermore, the direct binding of purified vimentin protein with LMWH was detected with surface plasmon resonance methods. However, when we used fluorescence-labeled heparin for 24 h to identify whether this binding occurred within cells, heparin was distributed principally around endothelial cells. Taken together, these findings suggest that the monoincubation of LMWH with HUVECs could inhibit PC-3M cell adhesion to, and migration through, endothelium. LMWH's regulation of vimentin plays a role in the antimetastatic action.