Department of Thoracic Oncology, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan.
Int J Oncol. 2010 Nov;37(5):1289-96. doi: 10.3892/ijo_00000780.
To study endothelial injury in vitro, we established a three-dimensional (3-D) blood vessel model in which human umbilical vein endothelial cells were grown in the presence of basic fibroblast growth factor and vascular endothelial growth factor. We then performed comparative studies on cisplatin (cis-platinum-diammine-dichloride, CDDP)-induced endothelial injury in 3-D and monolayer cultures. In 3-D culture, CDDP induced cell death and tube breakdown without DNA damage, whereas CDDP induced apoptosis accompanied by DNA damage in monolayer culture. CDDP also induced caspase-3 activation in a concentration-dependent manner in both cultures. A broad-spectrum caspase inhibitor, zVAD-fmk, failed to prevent CDDP-induced cell death and tube breakdown in 3-D culture, whereas zVAD-fmk suppressed CDDP-induced apoptosis in monolayer culture. A calpain inhibitor, MDL28170, attenuated CDDP-induced cell death and tube breakdown in 3-D culture, but not apoptosis in monolayer culture. These results showed that calpain is involved in CDDP-induced endothelial injury in 3-D culture and there are significant differences in signaling pathways between 3-D and monolayer cultures.
为了研究体外的内皮细胞损伤，我们建立了一个三维（3-D）血管模型，其中人脐静脉内皮细胞在碱性成纤维细胞生长因子和血管内皮生长因子的存在下生长。然后，我们对顺铂（顺铂二胺二氯，CDDP）在 3-D 和单层培养物中诱导的内皮损伤进行了比较研究。在 3-D 培养中，CDDP 诱导细胞死亡和管破裂，而不会引起 DNA 损伤，而 CDDP 在单层培养中诱导伴随 DNA 损伤的细胞凋亡。CDDP 还在两种培养物中以浓度依赖性方式诱导半胱天冬酶-3 的激活。广谱半胱天冬酶抑制剂 zVAD-fmk 未能阻止 3-D 培养物中 CDDP 诱导的细胞死亡和管破裂，但抑制了单层培养物中 CDDP 诱导的细胞凋亡。钙蛋白酶抑制剂 MDL28170 减轻了 3-D 培养物中 CDDP 诱导的细胞死亡和管破裂，但对单层培养物中的细胞凋亡没有影响。这些结果表明钙蛋白酶参与了 3-D 培养物中 CDDP 诱导的内皮细胞损伤，并且 3-D 和单层培养物之间存在信号通路的显著差异。
