Department of Urology, Urological Research Laboratory, Translational UroOncology, University Hospital Düsseldorf, Düsseldorf, Germany.
Methods Mol Biol. 2021;2195:99-111. doi: 10.1007/978-1-0716-0860-9_8.
Cisplatin resistance still remains a major obstacle in the standard chemotherapeutic approach in late-stage and metastatic testicular germ cell cancer (GCC) patients. This multifactorial and complex phenomenon arises (concomitantly) on several levels due to impaired transport, decreased adduct formation, increased DNA-repair, decreased apoptosis, or compensating pathways. Evaluation of novel therapeutic approaches and pharmacological inhibitors still remains necessary to treat cisplatin-resistant GCCs. In this chapter, we present in vitro techniques to measure cytotoxic impacts of chemotherapeutic drugs on GCC cell lines. Specifically, we will discuss the measurement of relative cell viability by XTT assay, as well as cell cycle distribution and apoptosis assay by Nicoletti- and Annexin V/PI apoptosis assay with subsequent flow cytometry, respectively, to evaluate the effects of cytotoxic treatment in GCC cell lines.
顺铂耐药仍然是晚期和转移性睾丸生殖细胞癌 (GCC) 患者标准化疗方法的主要障碍。这种多因素和复杂的现象是由于多种水平的转运受损、加合物形成减少、DNA 修复增加、细胞凋亡减少或补偿途径引起的。评估新的治疗方法和药理抑制剂仍然是治疗顺铂耐药 GCC 的必要手段。在这一章中，我们介绍了体外技术来测量化疗药物对 GCC 细胞系的细胞毒性影响。具体来说，我们将讨论通过 XTT 测定法测量相对细胞活力，以及通过尼科莱蒂法和 Annexin V/PI 凋亡测定法测量细胞周期分布和凋亡，分别通过流式细胞术进行，以评估细胞毒性治疗对 GCC 细胞系的影响。
