Monitoring drug efflux from sensitive and multidrug-resistant single cancer cells with microvoltammetry.
Monitoring drug efflux from sensitive and multidrug-resistant single cancer cells with microvoltammetry.
复制标题
使用微伏安法监测敏感和多重耐药单个癌细胞的药物流出。
DOI：
10.1021/ac9811773
复制
发表时间：
1999
影响因子：
7.4
通讯作者：
Gratzl,M
中科院分区：
文献类型：
--
作者：
Lu,H;Gratzl,M
多重耐药性（MDR）是某些癌细胞对一系列化学上不相关的药物最终产生的交叉耐药性。它归因于许多可能的生物物理过程，其中之一是耐药细胞的药物流出增加，导致细胞内药物积累和保留减少。在这项工作中，碳纤维微盘电极用于直接监测单个预装癌细胞的阿霉素流出。电化学清洁、吸附预浓缩以及环境氧气产生的电催化效应使得首次能够以非常接近（≤1μm）的良好时间分辨率（低至30秒/测量）检测到非常低的药物浓度（低至1nM），非常接近单个癌细胞。中国仓鼠卵巢癌细胞的敏感（AUXB1）和耐药（CHRC5）版本的结果表明，当两种细胞系预加载至相同的细胞内药物浓度时，耐药细胞表现出更高的初始流出率和更短的流出时间常数。这些观察结果与通过常规技术从相同细胞群获得的结果一致，证明微伏安法可用于监测单细胞水平的阿霉素流出。然而，与现有方法相比，其数据仅代表通常低时间分辨率下的平均细胞行为，此处描述的技术可以在高时间分辨率下的药物流出方面提供有关癌细胞群微观异质性的信息。由于直接在单个细胞上测量浓度，因此获得了流出的实际驱动力。这种方法可能会带来关于耐多药机制和前瞻性治疗的重要新信息。
Multidrug resistance (MDR) is the eventual cross-resistance of certain cancer cells to a series of chemically unrelated drugs. It is attributed to a number of possible biophysical processes, one of them being increased drug efflux from resistant cells which leads to a decreased intracellular drug accumulation and retention. In this work, a carbon fiber microdisk electrode was used to monitor directly doxorubicin efflux from single preloaded cancer cells. Electrochemical cleaning, adsorptive preconcentration, and an electrocatalytic effect due to ambient oxygen made it possible to detect eventually very low drug concentrations (down to 1 nM) at good temporal resolution (down to 30 s/measurement) very close (≤1 μm) to single cancer cells for the first time. The results from a sensitive (AUXB1) and a drug-resistant (CHRC5) version of Chinese hamster ovarian cancer cells show that resistant cells exhibit a much higher initial efflux rate and shorter efflux time constant when both cell lines are preloaded up to the same intracellular drug concentration. These observations are consistent with results obtained from populations of the same cells by conventional techniques, proving that microvoltammetry can be used to monitor doxorubicin efflux at the single-cell level. Compared with existing methodologies, however, whose data represent only average cell behavior at typically low temporal resolution, the technique described here can provide information on the microheterogeneity of cancer cell populations in terms of drug efflux at high temporal resolution. The actual driving force of efflux is obtained since concentrations are measured directly at individual cells. This approach may lead to important new information on the mechanisms and prospective treatments of MDR.