Laboratorio de Inmunoquímica I, Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N, Santo Tomás, Ciudad de Mexico 11340, Mexico.
Laboratorio de Bioingeniería de Tejidos, Departamento de Estudios de Posgrado e Investigación, Universidad Nacional Autónoma de México, Ciudad de Mexico 04360, Mexico.
Int J Mol Sci. 2024 May 9;25(10):5140. doi: 10.3390/ijms25105140.
Three-dimensional cell cultures have improved the evaluation of drugs for cancer therapy, due to their high similarity to solid tumors. In melanoma, autophagy appears to show a dual role depending on the progression of the disease. p62 protein has been proposed for the evaluation of autophagic flux since its expression is an indicator of the state of autophagy. Pentoxifylline (PTX) and Norcantharidin (NCTD) are drugs that have been shown to possess anticancer effects. In this work, we used B16F1 mouse melanoma cells in two-dimensional (2D) monolayer cultures and three-dimensional (3D) spheroids to test the effect of PTX and NCTD over the p62 expression. We analyzed the effect on p62 expression through Western blot and immunofluorescence assays. Our results indicate that PTX decreases p62 expression in both cell culture models, while Norcantharidin increases its expression in 3D cultures at 24 h. Therefore, these drugs could have a potential therapeutic use for the regulation of autophagy in melanoma, depending on the state of evolution of the disease.
三维细胞培养提高了癌症治疗药物的评估，因为它们与实体瘤非常相似。在黑色素瘤中，自噬似乎表现出双重作用，这取决于疾病的进展。p62 蛋白因其表达是自噬状态的指标，而被提议用于评估自噬流。己酮可可碱 (PTX) 和去甲斑蝥素 (NCTD) 是已被证明具有抗癌作用的药物。在这项工作中，我们使用 B16F1 小鼠黑色素瘤细胞在二维 (2D) 单层培养物和三维 (3D) 球体中测试 PTX 和 NCTD 对 p62 表达的影响。我们通过 Western blot 和免疫荧光分析来分析 p62 表达的影响。我们的结果表明，PTX 在两种细胞培养模型中均降低了 p62 的表达，而 NCTD 在 24 小时时增加了 3D 培养物中的表达。因此，这些药物可能具有调节黑色素瘤自噬的潜在治疗用途，具体取决于疾病的进化状态。
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