Rossini Elisa, Tamburello Mariangela, Abate Andrea, Zini Silvia, Ribaudo Giovanni, Gianoncelli Alessandra, Calza Stefano, Valcamonico Francesca, Suardi Nazareno R, Mirabella Giuseppe, Berruti Alfredo, Sigala Sandra
Section of Pharmacology, Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy.
Unit of Biostatistics and Bioinformatics, Department of Molecular and Translational Medicine, University of Brescia, 25123 Brescia, Italy.
Cells. 2024 Feb 20;13(5):368. doi: 10.3390/cells13050368.
Most patients with testicular germ cell tumors (GCTs) are treated with cisplatin (CP)-based chemotherapy. However, some of them may develop CP resistance and therefore represent a clinical challenge. Cyclin-dependent kinase 5 (CDK5) is involved in chemotherapy resistance in different types of cancer. Here, we investigated the possible role of CDK5 and other CDKs targeted by dinaciclib in nonseminoma cell models (both CP-sensitive and CP-resistant), evaluating the potential of the CDK inhibitor dinaciclib as a single/combined agent for the treatment of advanced/metastatic testicular cancer (TC).
The effects of dinaciclib and CP on sensitive and resistant NT2/D1 and NCCIT cell viability and proliferation were evaluated using MTT assays and direct count methods. Flow cytometry cell-cycle analysis was performed. The protein expression was assessed via Western blotting. The in vivo experiments were conducted in zebrafish embryos xenografted with TC cells.
Among all the CDKs analyzed, CDK5 protein expression was significantly higher in CP-resistant models. Dinaciclib reduced the cell viability and proliferation in each cell model, inducing changes in cell-cycle distribution. In drug combination experiments, dinaciclib enhances the CP effect both in vitro and in the zebrafish model.
Dinaciclib, when combined with CP, could be useful for improving nonseminoma TC response to CP.
大多数睾丸生殖细胞肿瘤（GCT）患者接受以顺铂（CP）为基础的化疗。然而，其中一些患者可能会产生CP耐药性，因此构成临床挑战。细胞周期蛋白依赖性激酶5（CDK5）参与不同类型癌症的化疗耐药。在此，我们研究了CDK5以及被dinaciclib靶向的其他CDK在非精原细胞瘤细胞模型（包括CP敏感和CP耐药模型）中的可能作用，评估CDK抑制剂dinaciclib作为单一/联合药物治疗晚期/转移性睾丸癌（TC）的潜力。
使用MTT法和直接计数法评估dinaciclib和CP对敏感及耐药的NT2/D1和NCCIT细胞活力和增殖的影响。进行流式细胞术细胞周期分析。通过蛋白质印迹法评估蛋白质表达。在移植了TC细胞的斑马鱼胚胎中进行体内实验。
在所有分析的CDK中，CDK5蛋白表达在CP耐药模型中显著更高。Dinaciclib降低了每个细胞模型中的细胞活力和增殖能力，诱导细胞周期分布发生变化。在药物联合实验中，dinaciclib在体外和斑马鱼模型中均增强了CP的作用。
Dinaciclib与CP联合使用，可能有助于提高非精原细胞瘤TC对CP的反应。
