Cell-based Drug and Health Products Development Research Unit, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Cell-based Drug and Health Products Development Research Unit, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand Department of Pharmacology and Physiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand
Anticancer Res. 2014 Dec;34(12):7111-7.
Resistance to chemotherapeutic agents, as well as enhanced metastasis, have been frequently reported in lung cancer.
Cytotoxicity and proliferative effects of cisplatin on H460 lung cancer cells were evaluated by the MTT assay. Migration capacity was evaluated by the wound healing assay. The number of filopodia per cell were detected by rhodamine-phalloidin staining assay. The changes of protein levels of integrins, and migration-related proteins in response to cisplatin at sub-toxic concentrations were determined by western blotting.
Herein we demonstrate for the first time that exposure to low concentrations of cisplatin results in increase of cell motility with the alteration of integrin expression. Cisplatin-treated cells exhibited a significant increase in the number of filopodia per cell in correlation with enhanced migration. Migration regulatory proteins, namely activated forms of focal-adhesion kinase (FAK) and ATP-dependent tyrosine kinase (AKT), were found to significantly be up-regulated in cisplatin-treated cells in comparison to those of the non-treated control. Active Rho A-GTP and Rac-GTP were found to be increased in accordance with activation of FAK/AKT signals. Furthermore, we found that such migration enhancement may be in part due to the integrin switch mediated by cisplatin treatment. Cisplatin induced a dramatic alteration in the integrin expression pattern by up-regulating integrin α4, αv, β1, and β5 which were previously reported to increase cell motility, while it had no effect on integrin α5, and β3.
As the integrin switch is a hallmark of highly aggressive cancer, these findings may provide insights for better understanding of cancer cell adaptation after exposure to cisplatin.
化疗药物耐药以及转移增强在肺癌中经常被报道。
通过 MTT 检测评估顺铂对 H460 肺癌细胞的细胞毒性和增殖作用。通过划痕愈合试验评估迁移能力。通过荧光鬼笔环肽染色试验检测细胞每根微丝的数量。通过 Western blot 检测亚毒性浓度顺铂作用下整合素和迁移相关蛋白的水平变化。
本文首次证明，低浓度顺铂暴露导致细胞迁移能力增加，同时改变整合素表达。与迁移增强相关，顺铂处理的细胞每根微丝的数量显著增加。与未经处理的对照组相比，迁移调节蛋白，即激活型粘着斑激酶（FAK）和 ATP 依赖型酪氨酸激酶（AKT），在顺铂处理的细胞中明显上调。发现活性 Rho A-GTP 和 Rac-GTP 与 FAK/AKT 信号的激活相一致而增加。此外，我们发现这种迁移增强可能部分归因于顺铂处理介导的整合素开关。顺铂通过上调先前报道能增加细胞迁移能力的整合素α4、αv、β1 和β5，显著改变整合素表达模式，而对整合素α5 和β3 没有影响。
由于整合素开关是高度侵袭性癌症的标志，这些发现可能为更好地理解顺铂暴露后癌细胞的适应提供新的见解。
