Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells

Maria De Salvo, Giovanna Maresca, Igea D'Agnano, Rodolfo Marchese, Antonio Stigliano, Raffaella Gagliassi, Ercole Brunetti, Giorgio H. Raza, Ugo De Paula, Barbara Bucci

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Purpose: We investigated the molecular mechanisms underlying the cytotoxic effect of Temozolomide (TMZ) in both O6-methylguanine-DNA methyl transferase (MGMT) depleted as well as undepleted glioblastoma cell lines. Since TMZ is used in clinics in combination with radiotherapy, we also studied the effects of TMZ in combination with ionising radiation (IR). Methods: Cell colony-forming ability was measured using a clonogenic assay. Cell cycle analysis and apoptosis were evaluated by Flow Cytometry (FCM). Proteins involved in cell cycle control were detected by Western blot and co-immunoprecipitation assays. Results: Our data showed that TMZ, independent of MGMT expression, inhibited glioblastoma cell growth via an irreversible G2 block in MGMT depleted cells or the induction of apoptosis in MGMT normal expressing cells. When TMZ was administered in combination with IR, apoptosis was greater than observed with either agent separately. This TMZ-induced apoptosis in the MGMT expressing cells occurred through Akt/Glycogen-Synthase-Kinase-3ß (GSK3ß) signalling and was mediated by Myelocytomatosis (c-Myc) oncoprotein. Indeed, TMZ phosphorylated/activated Akt led to phosphorylation/inactivation of GSK3ß which resulted in the stabilisation of c-Myc protein and subsequent modulation of the c-Myc target genes involved in the apoptotic processes. Conclusion: c-Myc expression could be considered a good indicator of TMZ effectiveness.

Original languageEnglish
Pages (from-to)518-533
Number of pages16
JournalInternational Journal of Radiation Biology
Volume87
Issue number5
DOIs
Publication statusPublished - May 2011

Fingerprint

temozolomide
Glioblastoma
Transferases
Apoptosis
DNA
Glycogen Synthase Kinase 3
Ionizing Radiation
Proto-Oncogene Proteins c-myc
myc Genes
Oncogene Proteins
Cell Cycle Checkpoints
Immunoprecipitation

Keywords

  • Akt/GSK3b pathway
  • apoptosis
  • c-Myc activation
  • glioblastoma cell lines
  • irradiation
  • temozolomide

ASJC Scopus subject areas

  • Radiology Nuclear Medicine and imaging
  • Radiological and Ultrasound Technology

Cite this

De Salvo, M., Maresca, G., D'Agnano, I., Marchese, R., Stigliano, A., Gagliassi, R., ... Bucci, B. (2011). Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells. International Journal of Radiation Biology, 87(5), 518-533. https://doi.org/10.3109/09553002.2011.556173

Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells. / De Salvo, Maria; Maresca, Giovanna; D'Agnano, Igea; Marchese, Rodolfo; Stigliano, Antonio; Gagliassi, Raffaella; Brunetti, Ercole; Raza, Giorgio H.; De Paula, Ugo; Bucci, Barbara.

In: International Journal of Radiation Biology, Vol. 87, No. 5, 05.2011, p. 518-533.

Research output: Contribution to journalArticle

De Salvo, M, Maresca, G, D'Agnano, I, Marchese, R, Stigliano, A, Gagliassi, R, Brunetti, E, Raza, GH, De Paula, U & Bucci, B 2011, 'Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells', International Journal of Radiation Biology, vol. 87, no. 5, pp. 518-533. https://doi.org/10.3109/09553002.2011.556173
De Salvo, Maria ; Maresca, Giovanna ; D'Agnano, Igea ; Marchese, Rodolfo ; Stigliano, Antonio ; Gagliassi, Raffaella ; Brunetti, Ercole ; Raza, Giorgio H. ; De Paula, Ugo ; Bucci, Barbara. / Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells. In: International Journal of Radiation Biology. 2011 ; Vol. 87, No. 5. pp. 518-533.
@article{3ce71334755f4a779930fcbac75a6cbb,
title = "Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells",
abstract = "Purpose: We investigated the molecular mechanisms underlying the cytotoxic effect of Temozolomide (TMZ) in both O6-methylguanine-DNA methyl transferase (MGMT) depleted as well as undepleted glioblastoma cell lines. Since TMZ is used in clinics in combination with radiotherapy, we also studied the effects of TMZ in combination with ionising radiation (IR). Methods: Cell colony-forming ability was measured using a clonogenic assay. Cell cycle analysis and apoptosis were evaluated by Flow Cytometry (FCM). Proteins involved in cell cycle control were detected by Western blot and co-immunoprecipitation assays. Results: Our data showed that TMZ, independent of MGMT expression, inhibited glioblastoma cell growth via an irreversible G2 block in MGMT depleted cells or the induction of apoptosis in MGMT normal expressing cells. When TMZ was administered in combination with IR, apoptosis was greater than observed with either agent separately. This TMZ-induced apoptosis in the MGMT expressing cells occurred through Akt/Glycogen-Synthase-Kinase-3{\ss} (GSK3{\ss}) signalling and was mediated by Myelocytomatosis (c-Myc) oncoprotein. Indeed, TMZ phosphorylated/activated Akt led to phosphorylation/inactivation of GSK3{\ss} which resulted in the stabilisation of c-Myc protein and subsequent modulation of the c-Myc target genes involved in the apoptotic processes. Conclusion: c-Myc expression could be considered a good indicator of TMZ effectiveness.",
keywords = "Akt/GSK3b pathway, apoptosis, c-Myc activation, glioblastoma cell lines, irradiation, temozolomide",
author = "{De Salvo}, Maria and Giovanna Maresca and Igea D'Agnano and Rodolfo Marchese and Antonio Stigliano and Raffaella Gagliassi and Ercole Brunetti and Raza, {Giorgio H.} and {De Paula}, Ugo and Barbara Bucci",
year = "2011",
month = "5",
doi = "10.3109/09553002.2011.556173",
language = "English",
volume = "87",
pages = "518--533",
journal = "International Journal of Radiation Biology",
issn = "0955-3002",
publisher = "Informa Healthcare",
number = "5",

}

TY - JOUR

T1 - Temozolomide induced c-Myc-mediated apoptosis via Akt signalling in MGMT expressing glioblastoma cells

AU - De Salvo, Maria

AU - Maresca, Giovanna

AU - D'Agnano, Igea

AU - Marchese, Rodolfo

AU - Stigliano, Antonio

AU - Gagliassi, Raffaella

AU - Brunetti, Ercole

AU - Raza, Giorgio H.

AU - De Paula, Ugo

AU - Bucci, Barbara

PY - 2011/5

Y1 - 2011/5

N2 - Purpose: We investigated the molecular mechanisms underlying the cytotoxic effect of Temozolomide (TMZ) in both O6-methylguanine-DNA methyl transferase (MGMT) depleted as well as undepleted glioblastoma cell lines. Since TMZ is used in clinics in combination with radiotherapy, we also studied the effects of TMZ in combination with ionising radiation (IR). Methods: Cell colony-forming ability was measured using a clonogenic assay. Cell cycle analysis and apoptosis were evaluated by Flow Cytometry (FCM). Proteins involved in cell cycle control were detected by Western blot and co-immunoprecipitation assays. Results: Our data showed that TMZ, independent of MGMT expression, inhibited glioblastoma cell growth via an irreversible G2 block in MGMT depleted cells or the induction of apoptosis in MGMT normal expressing cells. When TMZ was administered in combination with IR, apoptosis was greater than observed with either agent separately. This TMZ-induced apoptosis in the MGMT expressing cells occurred through Akt/Glycogen-Synthase-Kinase-3ß (GSK3ß) signalling and was mediated by Myelocytomatosis (c-Myc) oncoprotein. Indeed, TMZ phosphorylated/activated Akt led to phosphorylation/inactivation of GSK3ß which resulted in the stabilisation of c-Myc protein and subsequent modulation of the c-Myc target genes involved in the apoptotic processes. Conclusion: c-Myc expression could be considered a good indicator of TMZ effectiveness.

AB - Purpose: We investigated the molecular mechanisms underlying the cytotoxic effect of Temozolomide (TMZ) in both O6-methylguanine-DNA methyl transferase (MGMT) depleted as well as undepleted glioblastoma cell lines. Since TMZ is used in clinics in combination with radiotherapy, we also studied the effects of TMZ in combination with ionising radiation (IR). Methods: Cell colony-forming ability was measured using a clonogenic assay. Cell cycle analysis and apoptosis were evaluated by Flow Cytometry (FCM). Proteins involved in cell cycle control were detected by Western blot and co-immunoprecipitation assays. Results: Our data showed that TMZ, independent of MGMT expression, inhibited glioblastoma cell growth via an irreversible G2 block in MGMT depleted cells or the induction of apoptosis in MGMT normal expressing cells. When TMZ was administered in combination with IR, apoptosis was greater than observed with either agent separately. This TMZ-induced apoptosis in the MGMT expressing cells occurred through Akt/Glycogen-Synthase-Kinase-3ß (GSK3ß) signalling and was mediated by Myelocytomatosis (c-Myc) oncoprotein. Indeed, TMZ phosphorylated/activated Akt led to phosphorylation/inactivation of GSK3ß which resulted in the stabilisation of c-Myc protein and subsequent modulation of the c-Myc target genes involved in the apoptotic processes. Conclusion: c-Myc expression could be considered a good indicator of TMZ effectiveness.

KW - Akt/GSK3b pathway

KW - apoptosis

KW - c-Myc activation

KW - glioblastoma cell lines

KW - irradiation

KW - temozolomide

UR - http://www.scopus.com/inward/record.url?scp=79955602767&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79955602767&partnerID=8YFLogxK

U2 - 10.3109/09553002.2011.556173

DO - 10.3109/09553002.2011.556173

M3 - Article

C2 - 21405945

AN - SCOPUS:79955602767

VL - 87

SP - 518

EP - 533

JO - International Journal of Radiation Biology

JF - International Journal of Radiation Biology

SN - 0955-3002

IS - 5

ER -