Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66
Open Access
- 31 July 2020
- journal article
- research article
- Published by Frontiers Media SA in Frontiers in Physiology
- Vol. 11, 911
- https://doi.org/10.3389/fphys.2020.00911
Abstract
Introduction: Na+-K+-2Cl− cotransporter isoform 1 (NKCC1) is important in regulating intracellular K+ and Cl− homeostasis and cell volume. In this study, we investigated a role of NKCC1 in regulating glioma K+ influx and proliferation in response to apoptosis inducing chemotherapeutic drug temozolomide (TMZ). The efficacy of a new bumetanide (BMT)-derivative NKCC1 inhibitor STS66 [3-(butylamino)-2-phenoxy-5-[(2, 2, 2-trifluoroethylamino) methyl] benzenesulfonamide] in blocking NKCC1 activity was compared with well-established NKCC1 inhibitor BMT. Methods: NKCC1 activity in cultured mouse GL26 and SB28-GFP glioma cells was measured by Rb+ (K+) influx. The WNK1-SPAK/OSR1-NKCC1 signaling and AKT/ERK-mTOR signaling protein expression and activation were assessed by immunoblotting. Cell growth was determined by bromodeoxyuridine (BrdU) incorporation assay, MTT proliferation assay, and cell cycle analysis. Impact of STS66 and BMT on cell Rb+ influx and growth was measured in glioma cells treated with or without TMZ. Results: Rb+ influx assay showed that 10 μM BMT markedly decreased the total Rb+ influx and no additional inhibition detected at >10 μM BMT. In contrast, the maximum effects of STS66 on Rb+ influx inhibition were at 40–60 μM. Both BMT and STS66 reduced TMZ-mediated NKCC1 activation and protein upregulation. Glioma cell growth can be reduced by STS66. The most robust inhibition of glioma growth, cell cycle, and AKT/ERK signaling was achieved by the TMZ + STS66 treatment. Conclusion: The new BMT-derivative NKCC1 inhibitor STS66 is more effective than BMT in reducing glioma cell growth in part by inhibiting NKCC1-mediated K+ influx. TMZ + STS66 combination treatment reduces glioma cell growth via inhibiting cell cycle and AKT-ERK signaling.Funding Information
- National Natural Science Foundation of China (81801173)
- China Scholarship Council
This publication has 52 references indexed in Scilit:
- Hyperoxia resensitizes chemoresistant human glioblastoma cells to temozolomideJournal of Neuro-Oncology, 2012
- Regulation of Brain Tumor Dispersal by NKCC1 Through a Novel Role in Focal Adhesion RegulationPLoS Biology, 2012
- Inhibition of Na+-K+-2ClCotransporter isoform 1 Accelerates Temozolomidemediated Apoptosis in Glioblastoma Cancer CellsCellular Physiology and Biochemistry, 2012
- Hydrodynamic Cellular Volume Changes Enable Glioma Cell InvasionJournal of Neuroscience, 2011
- With-No-Lysine Kinase 3 (WNK3) stimulates glioma invasion by regulating cell volumeAmerican Journal of Physiology-Cell Physiology, 2011
- Ion channels and tranporters in cancer. 2. Ion channels and the control of cancer cell migrationAmerican Journal of Physiology-Cell Physiology, 2011
- Inhibition of the Sodium-Potassium-Chloride Cotransporter Isoform-1 Reduces Glioma InvasionCancer Research, 2010
- Chloride Accumulation Drives Volume Dynamics Underlying Cell Proliferation and MigrationJournal of Neurophysiology, 2009
- Physiology of Cell Volume Regulation in VertebratesPhysiological Reviews, 2009
- Cell shrinkage and monovalent cation fluxes: Role in apoptosisArchives of Biochemistry and Biophysics, 2007