Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66
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 temozolom...
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Frontiers Media S.A.
2020-07-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2020.00911/full |
| _version_ | 1818290340304519168 |
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| author | Lanxin Luo Lanxin Luo Jun Wang Jun Wang Dawei Ding Dawei Ding Md Nabiul Hasan Sung-Sen Yang Shih-Hua Lin Philipp Schreppel Baoshan Sun Yan Yin Thomas Erker Dandan Sun Dandan Sun |
| author_facet | Lanxin Luo Lanxin Luo Jun Wang Jun Wang Dawei Ding Dawei Ding Md Nabiul Hasan Sung-Sen Yang Shih-Hua Lin Philipp Schreppel Baoshan Sun Yan Yin Thomas Erker Dandan Sun Dandan Sun |
| author_sort | Lanxin Luo |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-13T02:26:37Z |
| format | Article |
| id | doaj.art-12fe751a292449c5b17969be452db028 |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-13T02:26:37Z |
| publishDate | 2020-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-12fe751a292449c5b17969be452db0282022-12-22T00:02:37ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2020-07-011110.3389/fphys.2020.00911561138Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66Lanxin Luo0Lanxin Luo1Jun Wang2Jun Wang3Dawei Ding4Dawei Ding5Md Nabiul Hasan6Sung-Sen Yang7Shih-Hua Lin8Philipp Schreppel9Baoshan Sun10Yan Yin11Thomas Erker12Dandan Sun13Dandan Sun14Department of Neurology, University of Pittsburgh, Pittsburgh, PA, United StatesDepartment of Neurosurgery, University of Minnesota, Minneapolis, MN, United StatesDepartment of Neurology, University of Pittsburgh, Pittsburgh, PA, United StatesDepartment of Neurology, The Second Hospital of Dalian Medical University, Dalian, ChinaDepartment of Neurology, University of Pittsburgh, Pittsburgh, PA, United StatesDepartment of Neurosurgery, University of Minnesota, Minneapolis, MN, United StatesDepartment of Neurology, University of Pittsburgh, Pittsburgh, PA, United StatesDivision of Nephrology, Department of Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, TaiwanDivision of Nephrology, Department of Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei, TaiwanDepartment of Medicinal Chemistry, University of Vienna, Vienna, AustriaPólo Dois Portos, Instituto National de Investigação Agrária e Veterinária, I.P., Quinta da Almoinha, Dois Portos, PortugalDepartment of Neurology, The Second Hospital of Dalian Medical University, Dalian, ChinaDepartment of Medicinal Chemistry, University of Vienna, Vienna, AustriaDepartment of Neurology, University of Pittsburgh, Pittsburgh, PA, United StatesVeterans Affairs Pittsburgh Health Care System, Geriatric Research, Educational and Clinical Center, Pittsburgh, PA, United StatesIntroduction: 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.https://www.frontiersin.org/article/10.3389/fphys.2020.00911/fullbumetanidebumetanide-derivative STS66cell volume regulationgliomaK+ influxNa+-K+-Cl− cotransporter |
| spellingShingle | Lanxin Luo Lanxin Luo Jun Wang Jun Wang Dawei Ding Dawei Ding Md Nabiul Hasan Sung-Sen Yang Shih-Hua Lin Philipp Schreppel Baoshan Sun Yan Yin Thomas Erker Dandan Sun Dandan Sun Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66 Frontiers in Physiology bumetanide bumetanide-derivative STS66 cell volume regulation glioma K+ influx Na+-K+-Cl− cotransporter |
| title | Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66 |
| title_full | Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66 |
| title_fullStr | Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66 |
| title_full_unstemmed | Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66 |
| title_short | Role of NKCC1 Activity in Glioma K+ Homeostasis and Cell Growth: New Insights With the Bumetanide-Derivative STS66 |
| title_sort | role of nkcc1 activity in glioma k homeostasis and cell growth new insights with the bumetanide derivative sts66 |
| topic | bumetanide bumetanide-derivative STS66 cell volume regulation glioma K+ influx Na+-K+-Cl− cotransporter |
| url | https://www.frontiersin.org/article/10.3389/fphys.2020.00911/full |
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