OncoPharma Research Group, Center for Health Technology and Services Research (CINTESIS), Rua Doutor Plácido da Costa, 4200-450 Porto, Portugal.
Faculty of Pharmacy, University of Porto, Rua Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal.
Biomolecules. 2022 Oct 16;12(10):1490. doi: 10.3390/biom12101490.
Drug combination and drug repurposing are two strategies that allow to find novel oncological therapies, in a faster and more economical process. In our previous studies, we developed a novel model of drug combination using antineoplastic and different repurposed drugs. We demonstrated the combinations of doxorubicin (DOX) + artesunate, DOX + chloroquine, paclitaxel (PTX) + fluoxetine, PTX + fluphenazine, and PTX + benztropine induce significant cytotoxicity in Michigan Cancer Foundation-7 (MCF-7) breast cancer cells. Furthermore, it was found that 5-FU + thioridazine and 5-fluorouracil (5-FU) + sertraline can synergistically induce a reduction in the viability of human colorectal adenocarcinoma cell line (HT-29). In this study, we aim to (1) evaluate the biosafety profile of these drug combinations for non-tumoral cells and (2) determine their mechanism of action in cancer cells. To do so, human fetal lung fibroblast cells (MRC-5) fibroblast cells were incubated for 48 h with all drugs, alone and in combination in concentrations of 0.25, 0.5, 1, 2, and 4 times their half-maximal inhibitory concentration (IC). Cell morphology and viability were evaluated. Next, we designed and constructed a cell microarray to perform immunohistochemistry studies for the evaluation of palmitoyl-protein thioesterase 1 (PPT1), Ki67, cleaved-poly (ADP-ribose) polymerase (cleaved-PARP), multidrug resistance-associated protein 2 (MRP2), P-glycoprotein (P-gp), and nuclear factor-kappa-B (NF-kB) p65 expression. We demonstrate that these combinations are cytotoxic for cancer cells and safe for non-tumoral cells at lower concentrations. Furthermore, it is also demonstrated that PPT1 may have an important role in the mechanism of action of these combinations, as demonstrated by their ability to decrease PPT1 expression. These results support the use of antimalarial and central nervous system (CNS) drugs in combination regimens with chemotherapeutic agents; nevertheless, additional studies are recommended to further explore their complete mechanisms of action.
药物联合和药物重定位是两种策略，可以在更快、更经济的过程中找到新的肿瘤治疗方法。在我们之前的研究中，我们开发了一种使用抗肿瘤药物和不同重定位药物的新的药物联合模型。我们证明了多柔比星（DOX）+青蒿琥酯、DOX+氯喹、紫杉醇（PTX）+氟西汀、PTX+氟奋乃静和 PTX+苯海索联合使用可显著诱导密歇根癌症基金会-7（MCF-7）乳腺癌细胞的细胞毒性。此外，还发现 5-氟尿嘧啶（5-FU）+噻氯匹定和 5-氟尿嘧啶（5-FU）+舍曲林可协同诱导人结直肠腺癌细胞系（HT-29）活力降低。在这项研究中，我们旨在（1）评估这些药物组合对非肿瘤细胞的生物安全性特征，（2）确定它们在癌细胞中的作用机制。为此，人胎儿肺成纤维细胞（MRC-5）成纤维细胞在浓度为其半最大抑制浓度（IC）的 0.25、0.5、1、2 和 4 倍的情况下与所有药物单独和联合孵育 48 小时，评估细胞形态和活力。接下来，我们设计并构建了细胞微阵列，以进行免疫组织化学研究，评估棕榈酰蛋白硫酯酶 1（PPT1）、Ki67、多聚（ADP-核糖）聚合酶（cleaved-PARP）、多药耐药相关蛋白 2（MRP2）、P-糖蛋白（P-gp）和核因子-κB（NF-κB）p65 的表达。我们证明，这些组合对癌细胞具有细胞毒性，在较低浓度下对非肿瘤细胞是安全的。此外，还证明 PPT1 可能在这些组合的作用机制中起重要作用，因为它们能够降低 PPT1 的表达。这些结果支持将抗疟药和中枢神经系统（CNS）药物与化疗药物联合使用，但建议进行更多研究以进一步探索其完整的作用机制。
