Rho GTPase protein expression and activation in murine monocytes/macrophages are not modulated by model biomaterial surfaces in serum-containing in vitro cultures
Rho GTPase protein expression and activation in murine monocytes/macrophages are not modulated by model biomaterial surfaces in serum-containing in vitro cultures
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DOI：
10.1163/156856206778530731
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发表时间：
2006-01-01
影响因子：
3.6
通讯作者：
Grainger, D. W.
中科院分区：
文献类型：
--
作者：
Godek, M. L.;Sampson, J. A.;Grainger, D. W.
通过检查模型生物材料上含血清培养物中的GT3表达和活化，研究了前单核细胞和（单核细胞-）巨噬细胞中的Rho GT3细胞信号传导级联。在生长于组织培养聚苯乙烯、聚苯乙烯、聚-L-丙交酯和Teflon AF表面上的细胞中测定Rho GDI和Rho GTP酶蛋白RhoA、Cdc 42和Rac 1的抑制。基于细胞成熟度（前单核细胞到单核细胞到巨噬细胞谱系）和模型表面化学比较蛋白质表达：Rho蛋白存在于模型表面上测试的大多数巨噬细胞中，表明Rho蛋白库可容易地用于响应于许多激活线索（包括生物材料表面接触）的信号传导事件。这些细胞系中的Rho GT3活化谱分别表明RAW 264.7中的活性Cdc 42和Rho蛋白、J774A.1中的Rac 1和Rho蛋白以及IC-21细胞系中的Cdc 42和Rac 1蛋白。总的来说，已知这些蛋白质在细胞粘附、扩散和运动所必需的所有基于肌动蛋白的细胞骨架重排中发挥关键作用，并且对于建立体内异物反应所需的细胞反应仍然很重要。基于细胞来源（原代与二次衍生细胞来源）或作为表面化学的函数的Rho GTdR蛋白表达水平的差异不显著。Rho GT3表达谱在前单核细胞非粘附前体细胞和成熟粘附单核细胞/巨噬细胞之间变化。在所有聚合物表面上从单核巨噬细胞系RAW 264.7和J774A.1检测到Rho GTdR蛋白的活性GTP结合形式，这表明虽然这些蛋白是细胞粘附行为的核心，但表面化学的差异不足以差异调节这些细胞类型中的GTdR活化。几天后，从在极性更大的组织培养聚苯乙烯和聚-L-丙交酯表面上培养的细胞中检测到活性Cdc 42，但在非极性聚苯乙烯和Teflon AF上生长的细胞中没有检测到活性Cdc 42，表明在含血清的培养物中对这种GTdR有一些表面影响。
The Rho GTPase cellular signaling cascade was investigated in pro-monocyte and (monocyte-)macrophage cells by examining GTPase expression and activation in serum-containing cultures on model biomaterials. Abundance of Rho GDI and the Rho GTPase proteins RhoA, Cdc42 and Rac1 was determined in cells grown on tissue-culture polystyrene, polystyrene, poly-L-lactide and Teflon (R) AF surfaces. Protein expression was compared based on cell maturity (promonocyte to monocyte to macrophage lineages) and by model surface chemistry: Rho proteins were present in the majority of macrophage cells tested on model surfaces suggesting that a pool of Rho proteins is readily available for signaling events in response to numerous activating cues, including biomaterials surface encounter. Rho GTPase activation profiles in these cell lines indicate active Cdc42 and Rho proteins in RAW 264.7, Rac1 and Rho in J774A.1, and Cdc42 and Rac1 in IC-21 cell lines, respectively. Collectively, these proteins are known to play critical roles in all actin-based cytoskeletal rearrangement necessary for cell adhesion, spreading and motility, and remain important to establishing cellular responses required for foreign body reactions in vivo. Differences in Rho GTPase protein expression levels based on cell sourcing (primary versus secondary-derived cell source), or as a function of surface chemistry were insignificant. Rho GTPase expression profiles varied between pro-monocytic non-adherent precursor cells and mature adherent monocyte/macrophage cells. The active GTP-bound forms of the Rho GTPase proteins were detected from monocyte-macrophage cell lines RAW 264.7 and J774A.1 on all polymer surfaces, suggesting that while these proteins are central to cell adhesive behavior, differences in surface chemistry are insufficient to differentially regulate GTPase activation in these cell types. Active Cdc42 was detected from cells cultured on the more-polar tissue-culture polystyrene and poly-L-lactide surfaces after several days, but absent from those grown on apolar polystyrene and Teflon (R) AF, indicating some surface influence on this GTPase in serum-containing cultures.