Ca(2+)‐activated K(+) channel K(Ca)1.1 as a therapeutic target to overcome chemoresistance in three‐dimensional sarcoma spheroid models

The large‐conductance Ca(2+)‐activated K(+) channel K(Ca)1.1 plays a pivotal role in tumor development and progression in several solid cancers. The three‐dimensional (3D) in vitro cell culture system is a powerful tool for cancer spheroid formation, and mimics in vivo solid tumor resistance to chemotherapy in the tumor microenvironment (TME). K(Ca)1.1 is functionally expressed in osteosarcoma and chondrosarcoma cell lines. K(Ca)1.1 activator‐induced hyperpolarizing responses were significantly larger in human osteosarcoma MG‐63 cells isolated from 3D spheroid models compared with in those from adherent 2D monolayer cells. The present study investigated the mechanisms underlying the upregulation of K(Ca)1.1 and its role in chemoresistance using a 3D spheroid model. K(Ca)1.1 protein expression levels were significantly elevated in the lipid‐raft‐enriched compartments of MG‐63 spheroids without changes in its transcriptional level. 3D spheroid formation downregulated the expression of the ubiquitin E3 ligase FBXW7, which is an essential contributor to K(Ca)1.1 protein degradation in breast cancer. The siRNA‐mediated inhibition of FBXW7 in MG‐63 cells from 2D monolayers upregulated K(Ca)1.1 protein expression. Furthermore, a treatment with a potent and selective K(Ca)1.1 inhibitor overcame the chemoresistance of the MG‐63 and human chondrosarcoma SW‐1353 spheroid models to paclitaxel, doxorubicin, and cisplatin. Among several multidrug resistance ATP‐binding cassette transporters, the expression of the multidrug resistance‐associated protein MRP1 was upregulated in both spheroids and restored by the inhibition of K(Ca)1.1. Therefore, the pharmacological inhibition of K(Ca)1.1 may be an attractive new strategy for acquiring resistance to chemotherapeutic drugs in the TME of K(Ca)1.1‐positive sarcomas.