title: In vitro evaluation of combined temozolomide and radiotherapy using X  rays and high-linear energy transfer radiation for glioblastoma.
abstract: High-linear energy transfer radiation offers superior biophysical properties over conventional radiotherapy and may have a great potential for treating radioresistant tumors, such as glioblastoma. However, very little pre-clinical data exists on the effects of high-LET radiation on glioblastoma cell lines and on the concomitant application of chemotherapy. This study investigates the in vitro effects of temozolomide in combination with low-energy protons and α particles. Cell survival, DNA damage and repair, and cell growth were examined in four human glioblastoma cell lines (LN18, T98G, U87 and U373) after treatment with either X rays, protons (LET 12.91 keV/μm), or α particles (LET 99.26 keV/μm) with or without concurrent temozolomide at clinically-relevant doses of 25 and 50 μM. The relative biological effectiveness at 10% survival (RBE(10)) increased as LET increased: 1.17 and 1.06 for protons, and 1.84 and 1.68 for α particles in the LN18 and U87 cell lines, respectively. Temozolomide administration increased cell killing in the O(6)-methylguanine DNA methyltransferase-methylated U87 and U373 cell lines. In contrast, temozolomide provided no therapeutic enhancement in the methylguanine DNA methyltransferase-unmethylated LN18 and T98G cell lines. In addition, the residual number of γ-H2AX foci at 24 h after treatment with radiation and concomitant temozolomide was found to be lower than or equal to that expected by DNA damage with either of the individual treatments. Kinetics of foci disappearance after X-ray and proton irradiation followed similar time courses; whereas, loss of γ-H2AX foci after α particle irradiation occurred at a slower rate than that by low-LET radiation (half-life 12.51-16.87 h). The combination of temozolomide with different radiation types causes additive rather than synergistic cytotoxicity. Nevertheless, particle therapy combined with chemotherapy may offer a promising alternative with the additional benefit of superior biophysical properties. It is also possible that new fractionation schedules could be designed to exploit the change in DNA repair kinetics when MGMT-methylated cells respond to high-LET radiation.
authors.affiliation:  Ion Beam Centre, Faculty of Engineering and Physical Sciences, University of Surrey, Guildford, Surrey GU2 7XH, United Kingdom. l.barazzuol@surrey.ac.uk
authors.forename: Lara
authors.initials: L
authors.lastname: Barazzuol
authors.forename: Raj
authors.initials: R
authors.lastname: Jena
authors.forename: Neil G
authors.initials: NG
authors.lastname: Burnet
authors.forename: Jonathan C G
authors.initials: JC
authors.lastname: Jeynes
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Merchant
authors.forename: Karen J
authors.initials: KJ
authors.lastname: Kirkby
authors.forename: Norman F
authors.initials: NF
authors.lastname: Kirkby
mesh_terms: D000512:Alpha Particles; D002455:Cell Division; D045744:Cell Line, Tumor / Q000187:drug effects / Q000528:radiation effects; D059248:Chemoradiotherapy; D019175:DNA Methylation / Q000187:drug effects; D015254:DNA Modification Methylases / Q000235:genetics / Q000378:metabolism; D045643:DNA Repair Enzymes / Q000235:genetics / Q000378:metabolism; D003606:Dacarbazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D005909:Glioblastoma / Q000473:pathology* / Q000628:therapy; D006801:Humans; D066298:In Vitro Techniques; D018499:Linear Energy Transfer; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D010315:Particle Accelerators; D017785:Photons; D011401:Promoter Regions, Genetic; D000077204:Temozolomide; D014410:Tumor Stem Cell Assay; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D009363:Neoplasm Proteins; D025521:Tumor Suppressor Proteins; D003606:Dacarbazine; D015254:DNA Modification Methylases; C515491:MGMT protein, human; D045643:DNA Repair Enzymes; D000077204:Temozolomide