Silveira Elaine, Cavalcante Isadora Pontes, Kremer Jean Lucas, de Mendonça Pedro Omori Ribeiro, Lotfi Claudimara Ferini Pacicco
Department of Anatomy, Institute of Biomedical Science, University of São Paulo, São Paulo, SP Brazil.
Cancer Cell Int. 2018 Mar 1;18:29. doi: 10.1186/s12935-018-0527-x. eCollection 2018.
New drugs for adrenocortical carcinoma (ACC) are needed because most patients undergo rapid disease progression despite surgery and adjuvant therapy with mitotane. In this study, we aimed to investigate the in vitro effects of different chemotherapy drugs, alone or combined with mitotane, on the viability of adrenocortical carcinoma cells.
Everolimus, sunitinib, zoledronic acid, imatinib and nilotinib cytotoxicity, alone or combined with mitotane were tested on ACC H295R cells in monolayer or spheroid cultures using MTS assays and confocal microscopy. Moreover, the nilotinib effects were investigated in spheroids cultured from patient tumor-derived ACC-T36 cells.
Morphological characterization of H295R cell spheroids using histochemistry was performed and showed that dense, homogenously sized, multicellular spheroids were obtained. We observed that sunitinib and nilotinib alone were equally effective in a monolayer preparation, whereas mitotane was the most effective even at a low dose. A combination of sunitinib and mitotane was the most effective treatment, with only 23.8% of cells in the monolayer remaining viable. Spheroid preparations showed resistance to different drugs, although the poor effect produced by mitotane alone was surprising, with a cell viability of 84.6% in comparison with 13.1% in monolayer cells. The most ineffective drugs in spheroid preparations were everolimus, zoledronic acid and imatinib. In both cell types, nilotinib, either alone or in combination with mitotane induced more significant cell viability inhibition in monolayer and spheroid preparations. In addition, the mechanism of nilotinib activity involves the ERK1/2 pathway.
Taken together, our data identified nilotinib as a cytotoxic drug that combined with ERK inhibitors deserves to be tested as a novel therapy for adrenocortical carcinoma.
肾上腺皮质癌（ACC）需要新的药物，因为尽管进行了手术和米托坦辅助治疗，但大多数患者的疾病仍会迅速进展。在本研究中，我们旨在研究不同化疗药物单独或与米托坦联合使用对肾上腺皮质癌细胞活力的体外影响。
使用MTS分析和共聚焦显微镜，在单层或球体培养的ACC H295R细胞中测试依维莫司、舒尼替尼、唑来膦酸、伊马替尼和尼洛替尼单独或与米托坦联合使用的细胞毒性。此外，在患者肿瘤来源的ACC-T36细胞培养的球体中研究了尼洛替尼的作用。
使用组织化学对H295R细胞球体进行了形态学表征，结果表明获得了致密、大小均匀的多细胞球体。我们观察到，在单层制剂中，舒尼替尼和尼洛替尼单独使用时效果相同，而米托坦即使在低剂量下也是最有效的。舒尼替尼和米托坦联合使用是最有效的治疗方法，单层中仅23.8%的细胞仍存活。球体制剂对不同药物表现出抗性，尽管单独使用米托坦产生的效果不佳令人惊讶，其细胞活力为84.6%，而单层细胞为13.1%。球体制剂中最无效的药物是依维莫司、唑来膦酸和伊马替尼。在两种细胞类型中，尼洛替尼单独或与米托坦联合使用在单层和球体制剂中均诱导更显著的细胞活力抑制。此外，尼洛替尼的活性机制涉及ERK1/2途径。
综上所述，我们的数据确定尼洛替尼为一种细胞毒性药物，与ERK抑制剂联合使用值得作为肾上腺皮质癌的新疗法进行测试。
