Department of Materials Science and Engineering, University of Delaware, Newark, DE 19716, USA.
Biomaterials. 2010 Jul;31(21):5700-18. doi: 10.1016/j.biomaterials.2010.03.017. Epub 2010 Apr 24.
Because prostate cancer cells metastasize to bone and exhibit osteoblastic features (osteomimicry), the interrelationships between bone-specific microenvironment and prostate cancer cells at sites of bone metastasis are critical to disease progression. In this work the bone marrow microenvironment in vitro was recreated both by tailoring scaffolds physical properties and by functionalizing electrospun polymer fibers with a bioactive peptide derived from domain IV of perlecan heparan sulfate proteoglycan. Electrospun poly (epsilon-caprolactone) (PCL) fibers and PCL/gelatin composite scaffolds were modified covalently with perlecan domain IV (PlnDIV) peptide. The expression of tight junction protein (E-cadherin) and focal adhesion kinase (FAK) phosphorylation on tyrosine 397 also were investigated. The described bioactive motif significantly enhanced adherence and infiltration of the metastatic prostate cancer cells on all modified electrospun substrates by day 5 post-seeding. Cells cultured on PlnDIV-modified matrices organized stress fibers and increased proliferation at statistically significant rates. Additional findings suggest that presence of PlnDIV peptide in the matrix reduced expression of tight junction protein and binding to PlnDIV peptide was accompanied by increased focal adhesion kinase (FAK) phosphorylation on tyrosine 397. We conclude that PlnDIV peptide supports key signaling events leading to proliferation, survival, and migration of C4-2B cancer cells; hence its incorporation into electrospun matrix is a key improvement to create a successful three-dimensional (3-D) pharmacokinetic cancer model.
由于前列腺癌细胞转移到骨骼并表现出成骨样特征（成骨模拟），因此骨骼特异性微环境与骨转移部位前列腺癌细胞之间的相互关系对疾病进展至关重要。在这项工作中，通过调整支架的物理性质和通过将源自硫酸乙酰肝素蛋白聚糖域 IV 的生物活性肽功能化来体外再现骨髓微环境。对聚（ε-己内酯）（PCL）纤维和 PCL/明胶复合支架进行共价修饰，以获得来自硫酸乙酰肝素蛋白聚糖域 IV 的肽（PlnDIV）。还研究了紧密连接蛋白（E-钙粘蛋白）的表达和粘着斑激酶（FAK）在酪氨酸 397 上的磷酸化。所述的生物活性基序显着增强了转移性前列腺癌细胞在所有修饰的静电纺丝基质上的粘附和渗透，在接种后第 5 天达到统计学显着水平。在 PlnDIV 修饰的基质上培养的细胞组织应力纤维并以统计学上显着的速度增加增殖。其他发现表明，基质中存在 PlnDIV 肽会降低紧密连接蛋白的表达，并且与 PlnDIV 肽的结合伴随着粘着斑激酶（FAK）在酪氨酸 397 上的磷酸化增加。我们得出结论，PlnDIV 肽支持导致 C4-2B 癌细胞增殖、存活和迁移的关键信号事件；因此，将其掺入静电纺丝基质是创建成功的三维（3-D）药代动力学癌症模型的关键改进。
