Towards multidrug resistance quantification using scanning electrochemical microscopy

癌细胞通过多药耐药性积极保护自己。 它在许多不同的癌症类型中的压倒性存在，包括急性白血病，结肠癌，肾癌，胰腺癌和类癌，严重破坏了当前化疗治疗的成功和患者生存率。 尽管这些治疗方法很有前途，而且往往不可替代，但它们中的许多都是低效的，因为它们针对的是难以量化的细胞耐药表型。因此，迫切需要开发具有改进的稳健性和灵敏度的新的生物分析方法，可以精确地量化与不同癌症类型相关的多药耐药性。因此，我们的研究计划致力于证明生物扫描电化学显微镜（SECM）可能是首选方法，因为它通过测量进出细胞的电化学探针的通量来评估细胞代谢，这是一种模拟耐药性的现象。为了令人信服，SECM测量必须在不同的细胞系（抗性和对照）上进行验证，这些细胞系以定义的模式共同固定在非导电（例如，塑料）支撑物。本发现基金的目的是对特异性过表达多药耐药蛋白MRP 1的不同人类癌细胞株进行SECM测量。在允许的五年时间表内，这将涉及设计一种软光刻方法，该方法能够将不同的细胞系（癌细胞和对照细胞）共同固定在塑料支持物上（例如，由Zeonor或聚苯乙烯制成），评估所选人肺癌和宫颈癌细胞系中MRP 1的表达水平，以及验证能够量化图案化癌细胞的抗性特征的通用电化学方案。

Cancer cells actively defend themselves through multidrug resistance. Its overwhelming presence in many different cancer types, including acute leukemia, colon, kidney, pancreas, and carcinoid cancers, seriously undermines the success of current chemotherapeutical treatments and patient survival rates. Although promising and often irreplaceable, many of these treatments are inefficient because they target a cellular resistance phenotype that is poorly quantified. It has therefore become urgent to develop new bioanalytical methods with improved robustness and sensitivity that can precisely quantify the multidrug resistance associated with different cancer types. As such, our research program is dedicated at demonstrating that biological scanning electrochemical microscopy (SECM) could be the method of choice since it evaluates cell metabolism by measuring the flux of electrochemical probes that are transported in and out of cells, a phenomena that mimics drug resistance. To be convincing, SECM measurements must be validated on different cell lines (resistant and control) that are co-immobilized in defined patterns on nonconductive (e.g., plastic) supports. The objective of the present discovery grant is to perform the SECM measurements on different strains of human cancer cells that specifically overexpress the multidrug resistant protein, MRP1. Within the allowed five-year timetable, this will involve the design of a soft-lithography method capable of co-immobilizing different cell lines (cancerous and control) on a plastic support (e.g., made of Zeonor or polystyrene), the evaluation of the expression levels of MRP1 in the chosen human lung and cervical cancer cell lines, and the validation of a general electrochemical protocol capable of quantifying the resistant signature of the patterned cancer cells.