Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026
In this paper we describe the influence of NU7026, a specific inhibitor of DNA-dependent protein kinase, phosphoinositide 3-kinase, and ATM-kinase on molecular and cellular mechanisms triggered by ionising irradiation in human T-lymphocyte leukaemic MOLT-4 cells. We studied the effect of this inhibi...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Karolinum Press
2012-01-01
|
| Series: | Acta Medica |
| Subjects: | |
| Online Access: | https://actamedica.lfhk.cuni.cz/55/2/0066/ |
| _version_ | 1829492832831799296 |
|---|---|
| author | Aleš Tichý Eva Novotná Kamila Ďurišová Barbora Šalovská Radka Sedlaříková Jaroslav Pejchal Lenka Zárybnická Jiřina Vávrová Zuzana Šinkorová Martina Řezáčová |
| author_facet | Aleš Tichý Eva Novotná Kamila Ďurišová Barbora Šalovská Radka Sedlaříková Jaroslav Pejchal Lenka Zárybnická Jiřina Vávrová Zuzana Šinkorová Martina Řezáčová |
| author_sort | Aleš Tichý |
| collection | DOAJ |
| description | In this paper we describe the influence of NU7026, a specific inhibitor of DNA-dependent protein kinase, phosphoinositide 3-kinase, and ATM-kinase on molecular and cellular mechanisms triggered by ionising irradiation in human T-lymphocyte leukaemic MOLT-4 cells. We studied the effect of this inhibitor (10 μM) combined with gammaradiation (1 Gy) leading to DNA damage response and induction of apoptosis. We used methods for apoptosis assessment (cell viability count and flow-cytometric analysis) and cell cycle analysis (DNA content measurement) and we detected expression and post-translational modifications (Western blotting) of proteins involved in DNA repair signalling pathways. Pre-treatment with NU7026 resulted into decreased activation of checkpoint kinase-2 (Thr68), p53 (Ser15 and Ser392), and histone H2A.X (Ser139) 2 hours after irradiation. Subsequently, combination of radiation and inhibitor led to decreased amount of cells in G2-phase arrest and into increased apoptosis after 72 hours. Our results indicate that in leukaemic cells the pre-incubation with inhibitor NU7026 followed by low doses of ionising radiation results in radio-sensitising of MOLT-4 cells via diminished DNA repair and delayed but pronounced apoptosis. This novel approach might offer new strategies in combined treatment of leukaemia diseases. |
| first_indexed | 2024-12-16T06:23:23Z |
| format | Article |
| id | doaj.art-863ee596f9ea4c379f25b2410648c158 |
| institution | Directory Open Access Journal |
| issn | 1211-4286 1805-9694 |
| language | English |
| last_indexed | 2024-12-16T06:23:23Z |
| publishDate | 2012-01-01 |
| publisher | Karolinum Press |
| record_format | Article |
| series | Acta Medica |
| spelling | doaj.art-863ee596f9ea4c379f25b2410648c1582022-12-21T22:41:05ZengKarolinum PressActa Medica1211-42861805-96942012-01-01552667310.14712/18059694.2015.57Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026Aleš Tichý0Eva Novotná1Kamila Ďurišová2Barbora Šalovská3Radka Sedlaříková4Jaroslav Pejchal5Lenka Zárybnická6Jiřina Vávrová7Zuzana Šinkorová8Martina Řezáčová9Charles University in Prague, Faculty of Medicine in Hradec Králové, Czech Republic: Institute of Medical Biochemistry; University of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyCharles University in Prague, Faculty of Medicine in Hradec Králové, Czech Republic: Institute of Medical Biochemistry; University of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Centrum of Advanced StudiesUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyUniversity of Defence in Brno, Faculty of Health Sciences in Hradec Králové, Czech Republic: Department of RadiobiologyCharles University in Prague, Faculty of Medicine in Hradec Králové, Czech Republic: Institute of Medical BiochemistryIn this paper we describe the influence of NU7026, a specific inhibitor of DNA-dependent protein kinase, phosphoinositide 3-kinase, and ATM-kinase on molecular and cellular mechanisms triggered by ionising irradiation in human T-lymphocyte leukaemic MOLT-4 cells. We studied the effect of this inhibitor (10 μM) combined with gammaradiation (1 Gy) leading to DNA damage response and induction of apoptosis. We used methods for apoptosis assessment (cell viability count and flow-cytometric analysis) and cell cycle analysis (DNA content measurement) and we detected expression and post-translational modifications (Western blotting) of proteins involved in DNA repair signalling pathways. Pre-treatment with NU7026 resulted into decreased activation of checkpoint kinase-2 (Thr68), p53 (Ser15 and Ser392), and histone H2A.X (Ser139) 2 hours after irradiation. Subsequently, combination of radiation and inhibitor led to decreased amount of cells in G2-phase arrest and into increased apoptosis after 72 hours. Our results indicate that in leukaemic cells the pre-incubation with inhibitor NU7026 followed by low doses of ionising radiation results in radio-sensitising of MOLT-4 cells via diminished DNA repair and delayed but pronounced apoptosis. This novel approach might offer new strategies in combined treatment of leukaemia diseases.https://actamedica.lfhk.cuni.cz/55/2/0066/Inhibitor NU7026DNA-PKATMPI3-KIonizing radiationLeukaemia |
| spellingShingle | Aleš Tichý Eva Novotná Kamila Ďurišová Barbora Šalovská Radka Sedlaříková Jaroslav Pejchal Lenka Zárybnická Jiřina Vávrová Zuzana Šinkorová Martina Řezáčová Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026 Acta Medica Inhibitor NU7026 DNA-PK ATM PI3-K Ionizing radiation Leukaemia |
| title | Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026 |
| title_full | Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026 |
| title_fullStr | Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026 |
| title_full_unstemmed | Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026 |
| title_short | Radio-Sensitization of Human Leukaemic MOLT-4 Cells by DNA-Dependent Protein Kinase Inhibitor, NU7026 |
| title_sort | radio sensitization of human leukaemic molt 4 cells by dna dependent protein kinase inhibitor nu7026 |
| topic | Inhibitor NU7026 DNA-PK ATM PI3-K Ionizing radiation Leukaemia |
| url | https://actamedica.lfhk.cuni.cz/55/2/0066/ |
| work_keys_str_mv | AT alestichy radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT evanovotna radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT kamiladurisova radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT barborasalovska radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT radkasedlarikova radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT jaroslavpejchal radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT lenkazarybnicka radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT jirinavavrova radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT zuzanasinkorova radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 AT martinarezacova radiosensitizationofhumanleukaemicmolt4cellsbydnadependentproteinkinaseinhibitornu7026 |