Madrid Maria Fernanda, Mendoza Eleicy Nathaly, Padilla Ana Lizeth, Choquenaira-Quispe Celia, de Jesus Guimarães Celina, de Melo Pereira João Victor, Barros-Nepomuceno Francisco Washington Araújo, Lopes Dos Santos Ingredy, Pessoa Claudia, de Moraes Filho Manoel Odorico, Rocha Danilo Damasceno, Ferreira Paulo Michel Pinheiro
Drug Research and Development Center (NPDM), Department of Physiology and Pharmacology, Federal University of Ceará, Fortaleza, Brazil.
Pharmaceutical Sciences, Faculty of Pharmacy, Dentistry, and Nursing, Federal University of Ceará, Fortaleza, Brazil.
J Toxicol Environ Health B Crit Rev. 2025 Jan 2;28(1):1-27. doi: 10.1080/10937404.2024.2407452. Epub 2024 Oct 3.
The overexpression of ATP-binding cassette (ABC) transporters contributes to the failure of chemotherapies and symbolizes a great challenge in oncology, associated with the adaptation of tumor cells to anticancer drugs such that these transporters become less effective, a mechanism known as multidrug resistance (MDR). The aim of this review is to present the most widely used methodologies for induction and comprehension of models for detection of multidrug-resistant (MDR) modulators or inhibitors, including biochemical and morphological techniques for chemosensitivity studies. The overexpression of MDR proteins, predominantly, the subfamily glycoprotein-1 (P-gp or ABCB1) multidrug resistance, multidrug resistance-associated protein 1 (MRP1 or ABCCC1), multidrug resistance-associated protein 2 (MRP2 or ABCC2) and cancer resistance protein (ABCG2), in chemotherapy-exposed cancer lines have been established/investigated by several techniques. Amongst these techniques, the most used are (i) colorimetric/fluorescent indirect bioassays, (ii) rhodamine and efflux analysis, (iii) release of 3,30-diethyloxacarbocyanine iodide by fluorescence microscopy and flow cytometry to measure P-gp function and other ABC transporters, (iv) exclusion of calcein-acetoxymethylester, (v) ATPase assays to distinguish types of interaction with ABC transporters, (vi) morphology to detail phenotypic characteristics in transformed cells, (vii) molecular testing of resistance-related proteins (RT-qPCR) and (viii) 2D and 3D models, (ix) organoids, and (x) microfluidic technology. Then, models for detecting chemotherapy MDR cells to assess innovative therapies to modulate or inhibit tumor cell growth and overcome clinical resistance. It is noteworthy that different therapies including anti-miRNAs, antibody-drug conjugates (to natural products), and epigenetic modifications were also considered as promising alternatives, since currently no anti-MDR therapies are able to improve patient quality of life. Therefore, there is also urgency for new clinical markers of resistance to more reliably reflect effectiveness of novel antitumor drugs.
ATP结合盒（ABC）转运蛋白的过表达导致化疗失败，是肿瘤学领域的一大挑战，这与肿瘤细胞对抗癌药物的适应性有关，使得这些转运蛋白的效果降低，这种机制称为多药耐药（MDR）。本综述的目的是介绍诱导和理解多药耐药（MDR）调节剂或抑制剂检测模型最广泛使用的方法，包括用于化学敏感性研究的生化和形态学技术。化疗暴露的癌症细胞系中MDR蛋白的过表达，主要是糖蛋白-1（P-gp或ABCB1）多药耐药亚家族、多药耐药相关蛋白1（MRP1或ABCC1）、多药耐药相关蛋白2（MRP2或ABCC2）和抗癌蛋白（ABCG2），已通过多种技术得以确立/研究。在这些技术中，最常用的有：（i）比色/荧光间接生物测定法；（ii）罗丹明和外排分析；（iii）通过荧光显微镜和流式细胞术检测3,3'-二乙基氧杂羰花青碘化物的释放以测量P-gp功能和其他ABC转运蛋白；（iv）钙黄绿素-乙酰氧基甲酯排除法；（v）ATP酶测定以区分与ABC转运蛋白的相互作用类型；（vi）形态学以详细描述转化细胞的表型特征；（vii）耐药相关蛋白的分子检测（RT-qPCR）；（viii）二维和三维模型；（ix）类器官；以及（x）微流控技术。然后，建立检测化疗MDR细胞的模型，以评估调节或抑制肿瘤细胞生长并克服临床耐药性的创新疗法。值得注意的是，包括抗miRNA、抗体-药物偶联物（针对天然产物）和表观遗传修饰在内的不同疗法也被视为有前景的替代方案，因为目前尚无抗MDR疗法能够改善患者生活质量。因此，也迫切需要新的耐药临床标志物，以更可靠地反映新型抗肿瘤药物的疗效。
