Laboratorio Nacional de Canalopatías, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior s/n, C.U. 04510, Mexico City, Mexico.
Departamento de Neurociencia Cognitiva, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Circuito Exterior s/n, C.U. 04510, Mexico City, Mexico.
Pharmacol Rep. 2021 Dec;73(6):1744-1753. doi: 10.1007/s43440-021-00304-5. Epub 2021 Jul 2.
Ion channels have been proposed as therapeutic targets for different types of malignancies. One of the most studied ion channels in cancer is the voltage-gated potassium channel ether-à-go-go 1 or Kv10.1. Various studies have shown that Kv10.1 expression induces the proliferation of several cancer cell lines and in vivo tumor models, while blocking or silencing inhibits proliferation. Kv10.1 is a promising target for drug discovery modulators that could be used in cancer treatment. This work aimed to screen for new Kv10.1 channel modulators using a thallium influx-based assay.
Pharmacological effects of small molecules on Kv10.1 channel activity were studied using a thallium-based fluorescent assay and patch-clamp electrophysiological recordings, both performed in HEK293 stably expressing the human Kv10.1 potassium channel.
In thallium-sensitive fluorescent assays, we found that the small molecules loperamide and amitriptyline exert a potent inhibition on the activity of the oncogenic potassium channel Kv10.1. These results were confirmed by electrophysiological recordings, which showed that loperamide and amitriptyline decreased the amplitude of Kv10.1 currents in a dose-dependent manner. Both drugs could be promising tools for further studies.
Thallium-sensitive fluorescent assay represents a reliable methodological tool for the primary screening of different molecules with potential activity on Kv10.1 channels or other K channels.
离子通道已被提议作为治疗不同类型恶性肿瘤的靶点。在癌症中研究最多的离子通道之一是电压门控钾通道醚-α--go-go 1 或 Kv10.1。多项研究表明，Kv10.1 的表达可诱导多种癌细胞系和体内肿瘤模型的增殖，而阻断或沉默则可抑制增殖。Kv10.1 是一种有前途的药物发现调节剂靶点，可用于癌症治疗。这项工作旨在使用基于铊内流的测定法筛选新的 Kv10.1 通道调节剂。
使用基于铊的荧光测定法和膜片钳电生理记录研究小分子对 Kv10.1 通道活性的药理学作用，这两种方法均在稳定表达人 Kv10.1 钾通道的 HEK293 中进行。
在铊敏感的荧光测定中，我们发现小分子洛哌丁胺和阿米替林对致癌钾通道 Kv10.1 的活性具有强大的抑制作用。电生理记录证实了这一结果，表明洛哌丁胺和阿米替林以剂量依赖性方式降低 Kv10.1 电流的幅度。这两种药物都可能是进一步研究的有前途的工具。
铊敏感荧光测定法是一种可靠的方法学工具，可用于对 Kv10.1 通道或其他 K 通道具有潜在活性的不同分子进行初步筛选。
