Doxorubicin (Doxo) is the most effective chemotherapeutic agent for the treatment of breast cancer. However, resistance to Doxo is common. Adjuvant compounds capable of modulating mechanisms involved in Doxo resistance may potentiate the effectiveness of the drug. Resveratrol (Rsv) has been tested as an adjuvant in mammary malignancies. However, the cellular and molecular mechanisms underlying the effects of cotreatment with Doxo and Rsv in breast cancer are poorly understood. Here, we combined in vitro and in silico analysis to characterize these mechanisms. In vitro, we employed a clinically relevant experimental design consisting of acute (24 h) treatment followed by 15 days of analysis. Acute Rsv potentiated the long-lasting effect of Doxo through the induction of apoptosis and senescence. Cells that survived to the cotreatment triggered high levels of autophagy. Autophagy inhibition during its peak of activation but not concomitant with Doxo+Rsv increased the long-term toxicity of the cotreatment. To uncover key proteins potentially associated with in vitro effects, an in silico multistep strategy was implemented. Chemical-protein networks were predicted based on constitutive gene expression of MCF7 cells and interatomic data from breast cancer. Topological analysis, KM survival analysis, and a quantitative model based on the connectivity between apoptosis, senescence, and autophagy were performed. We found seven putative genes predicted to be modulated by Rsv in the context of Doxo treatment: CCND1, CDH1, ESR1, HSP90AA1, MAPK3, PTPN11, and RPS6KB1. Six out of these seven genes have been experimentally proven to be modulated by Rsv in cancer cells, with 4 of the 6 genes in MCF7 cells. In conclusion, acute Rsv potentiated the long-term toxicity of Doxo in breast cancer potentially through the modulation of genes and mechanisms involved in Doxo resistance. Rational autophagy inhibition potentiated the effects of Rsv+Doxo, a strategy that should be further tested in animal models.

中文翻译：

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白藜芦醇和阿霉素联合治疗乳腺癌所触发的细胞机制：转化体外计算机模型

阿霉素（Doxo）是治疗乳腺癌最有效的化疗药物。然而，对 Doxo 的耐药性很常见。能够调节 Doxo 耐药机制的佐剂化合物可能会增强药物的有效性。白藜芦醇 (Rsv) 已被测试作为乳腺恶性肿瘤的佐剂。然而，Doxo 和 Rsv 联合治疗乳腺癌的细胞和分子机制尚不清楚。在这里，我们结合体外和计算机分析来表征这些机制。在体外，我们采用了临床相关的实验设计，包括急性（24 小时）治疗和随后 15 天的分析。Acute Rsv 通过诱导细胞凋亡和衰老来增强 Doxo 的长期作用。共处理后存活的细胞引发了高水平的自噬。在其激活高峰期间的自噬抑制，但不与 Doxo+Rsv 同时进行，会增加联合治疗的长期毒性。为了发现可能与体外效应相关的关键蛋白质，实施了计算机模拟多步骤策略。根据 MCF7 细胞的组成型基因表达和乳腺癌的原子间数据预测化学蛋白质网络。进行了拓扑分析、KM 存活分析以及基于细胞凋亡、衰老和自噬之间连接的定量模型。我们发现了 7 个预计在 Doxo 治疗背景下受 Rsv 调节的基因：CCND1、CDH1、ESR1、HSP90AA1、MAPK3、PTPN11 和 RPS6KB1。这七个基因中的六个已被实验证明在癌细胞中受到 Rsv 的调节，其中 6 个基因中的 4 个在 MCF7 细胞中。总之，急性 Rsv 可能通过调节与 Doxo 耐药相关的基因和机制，增强了 Doxo 对乳腺癌的长期毒性。合理的自噬抑制增强了 Rsv+Doxo 的效果，这一策略应该在动物模型中进一步测试。