The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures
The hEag1 (Kv10.1) K<sup>+</sup> channel is normally found in the brain, but it is ectopically expressed in tumor cells, including osteosarcoma. Based on the pivotal role of ion channels in osteogenesis, we tested whether pharmacological modulation of hEag1 may affect osteogenic differen...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/18/10533 |
| _version_ | 1797487344243179520 |
|---|---|
| author | Beáta Mészáros Agota Csoti Tibor G. Szanto Andrea Telek Katalin Kovács Agnes Toth Julianna Volkó Gyorgy Panyi |
| author_facet | Beáta Mészáros Agota Csoti Tibor G. Szanto Andrea Telek Katalin Kovács Agnes Toth Julianna Volkó Gyorgy Panyi |
| author_sort | Beáta Mészáros |
| collection | DOAJ |
| description | The hEag1 (Kv10.1) K<sup>+</sup> channel is normally found in the brain, but it is ectopically expressed in tumor cells, including osteosarcoma. Based on the pivotal role of ion channels in osteogenesis, we tested whether pharmacological modulation of hEag1 may affect osteogenic differentiation of osteosarcoma cell lines. Using molecular biology (RT-PCR), electrophysiology (patch-clamp) and pharmacology (astemizole sensitivity, IC<sub>50</sub> = 0.135 μM) we demonstrated that SaOS-2 osteosarcoma cells also express hEag1 channels. SaOS-2 cells also express to KCa1.1 K<sup>+</sup> channels as shown by mRNA expression and paxilline sensitivity of the current. The inhibition of hEag1 (2 μM astemizole) or KCa1.1 (1 mM TEA) alone did not induce Ca<sup>2+</sup> deposition in SaOS-2 cultures, however, these inhibitors, at identical concentrations, increased Ca<sup>2+</sup> deposition evoked by the classical or pathological (inorganic phosphate, Pi) induction pathway without causing cytotoxicity, as reported by three completer assays (LDH release, MTT assay and SRB protein assay). We observed a similar effect of astemizole on Ca<sup>2+</sup> deposition in MG-63 osteosarcoma cultures as well. We propose that the increase in the osteogenic stimuli-induced mineral matrix formation of osteosarcoma cell lines by inhibiting hEag1 may be a useful tool to drive terminal differentiation of osteosarcoma. |
| first_indexed | 2024-03-09T23:46:18Z |
| format | Article |
| id | doaj.art-f21ecadb81d0467f81fe8e6581bf43d6 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:46:18Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f21ecadb81d0467f81fe8e6581bf43d62023-11-23T16:43:24ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-09-0123181053310.3390/ijms231810533The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma CulturesBeáta Mészáros0Agota Csoti1Tibor G. Szanto2Andrea Telek3Katalin Kovács4Agnes Toth5Julianna Volkó6Gyorgy Panyi7Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Medical Chemistry, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryDepartment of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Life Science Building, Egyetem Ter 1, H-4032 Debrecen, HungaryThe hEag1 (Kv10.1) K<sup>+</sup> channel is normally found in the brain, but it is ectopically expressed in tumor cells, including osteosarcoma. Based on the pivotal role of ion channels in osteogenesis, we tested whether pharmacological modulation of hEag1 may affect osteogenic differentiation of osteosarcoma cell lines. Using molecular biology (RT-PCR), electrophysiology (patch-clamp) and pharmacology (astemizole sensitivity, IC<sub>50</sub> = 0.135 μM) we demonstrated that SaOS-2 osteosarcoma cells also express hEag1 channels. SaOS-2 cells also express to KCa1.1 K<sup>+</sup> channels as shown by mRNA expression and paxilline sensitivity of the current. The inhibition of hEag1 (2 μM astemizole) or KCa1.1 (1 mM TEA) alone did not induce Ca<sup>2+</sup> deposition in SaOS-2 cultures, however, these inhibitors, at identical concentrations, increased Ca<sup>2+</sup> deposition evoked by the classical or pathological (inorganic phosphate, Pi) induction pathway without causing cytotoxicity, as reported by three completer assays (LDH release, MTT assay and SRB protein assay). We observed a similar effect of astemizole on Ca<sup>2+</sup> deposition in MG-63 osteosarcoma cultures as well. We propose that the increase in the osteogenic stimuli-induced mineral matrix formation of osteosarcoma cell lines by inhibiting hEag1 may be a useful tool to drive terminal differentiation of osteosarcoma.https://www.mdpi.com/1422-0067/23/18/10533hEag1 potassium channelSaOS-2 osteosarcoma cellsCa<sup>2+</sup> depositionKv10.1 |
| spellingShingle | Beáta Mészáros Agota Csoti Tibor G. Szanto Andrea Telek Katalin Kovács Agnes Toth Julianna Volkó Gyorgy Panyi The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures International Journal of Molecular Sciences hEag1 potassium channel SaOS-2 osteosarcoma cells Ca<sup>2+</sup> deposition Kv10.1 |
| title | The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures |
| title_full | The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures |
| title_fullStr | The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures |
| title_full_unstemmed | The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures |
| title_short | The hEag1 K<sup>+</sup> Channel Inhibitor Astemizole Stimulates Ca<sup>2+</sup> Deposition in SaOS-2 and MG-63 Osteosarcoma Cultures |
| title_sort | heag1 k sup sup channel inhibitor astemizole stimulates ca sup 2 sup deposition in saos 2 and mg 63 osteosarcoma cultures |
| topic | hEag1 potassium channel SaOS-2 osteosarcoma cells Ca<sup>2+</sup> deposition Kv10.1 |
| url | https://www.mdpi.com/1422-0067/23/18/10533 |
| work_keys_str_mv | AT beatameszaros theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT agotacsoti theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT tiborgszanto theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT andreatelek theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT katalinkovacs theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT agnestoth theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT juliannavolko theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT gyorgypanyi theheag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT beatameszaros heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT agotacsoti heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT tiborgszanto heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT andreatelek heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT katalinkovacs heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT agnestoth heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT juliannavolko heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures AT gyorgypanyi heag1ksupsupchannelinhibitorastemizolestimulatescasup2supdepositioninsaos2andmg63osteosarcomacultures |