title: The effects of pentoxifylline on the survival of human glioma cells with continuous and intermittent stereotactic radiosurgery irradiation.
abstract: OBJECTIVE
  In linac-based stereotactic radiosurgery, treatment is delivered intermittently via multiple individual small radiotherapy arcs. The time lapses between the individual arcs permit greater damage repair and increased tumor cell survival in comparison with continuous irradiation. Because pentoxifylline (PTX) has been reported to prevent radiation-induced cell cycle arrest at the G2/M checkpoint, where damage repair is critically linked to cell survival, we hypothesized that PTX would exert a favorable radiosensitization effect by reducing the recovery observed during intermittent stereotactic radiosurgery.
  
  
  METHODS
  The human glioma cell line T98G was used to study the effects of continuous vs. intermittent irradiation with or without PTX. Cell cycle patterns were studied using flow cytometry. Clonogenic assays of single cells and spheroid outgrowth assays provided a quantitative measure of PTX-mediated radiosensitization. The PTX effect upon cells in low oxygen conditions was also studied in vitro after enzymatic oxygen scavenging.
  
  
  RESULTS
  Flow Cytometry: T98G cells exposed to both continuous and intermittent irradiation exhibit similar arrest at the G2/M checkpoint. The addition of 2 mM PTX significantly reduced the radiation-induced G2/M block in both irradiation schemes. Clonogenic Assays: The same PTX concentration applied before a continuous dose of 12 Gy, two intermittent doses of 6 Gy, or three intermittent doses of 4 Gy, all given within a 1-h interval, consistently caused radiosensitization. The drug enhancement ratios for PTX were 1.5, 2.7, and 6.0 for the continuous and two different intermittent dose schedules, respectively. Adding PTX after irradiation yielded lower enhancement ratios than pre-irradiation application. A similar pattern was observed after total doses of 4, 6, 9, or 12 Gy, as well. In low oxygen conditions, PTX was seen to have the same effects as in normoxic conditions. Spheroid Outgrowth Assays: The in vitro PTX effects were replicated in the spheroid outgrowth assays.
  
  
  CONCLUSIONS
  In human glioma cells, PTX abrogates the radiation-induced G2/M block observed after either continuous radiation exposure or intermittent exposures modeling clinical linac-based radiosurgery. The PTX-mediated reduction of the G2/M block translates into radiosensitization, most notably during intermittent exposures, and is presumably a consequence of diminished DNA damage repair at the G2/M checkpoint, though other contributing effects cannot be ruled out. The radiosensitization effect of PTX is sustained under low oxygen conditions. These results support consideration of the clinical evaluation of PTX to enhance the efficacy of linac-based radiosurgery involving intermittent irradiation through multiple arcs.
authors.affiliation: Department of Radiation Oncology, Medical College of Virginia Hospitals of Virginia Commonwealth University, Richmond, VA 23298-0058, USA. kdwillia@vcu.edu
authors.forename: Kerry W
authors.initials: KW
authors.lastname: Eley
authors.forename: Stanley H
authors.initials: SH
authors.lastname: Benedict
authors.forename: Theodore D K
authors.initials: TD
authors.lastname: Chung
authors.forename: Brian D
authors.initials: BD
authors.lastname: Kavanagh
authors.forename: William C
authors.initials: WC
authors.lastname: Broaddus
authors.forename: Rupert K A
authors.initials: RK
authors.lastname: Schmidt-Ullrich
authors.forename: Peck-Sun
authors.initials: PS
authors.lastname: Lin
mesh_terms: D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D004260:DNA Repair; D005434:Flow Cytometry / Q000379:methods; D016195:G2 Phase / Q000187:drug effects* / Q000528:radiation effects; D005910:Glioma / Q000473:pathology / Q000601:surgery*; D006801:Humans; D010431:Pentoxifylline / Q000494:pharmacology*; D011829:Radiation Dosage; D011837:Radiation-Protective Agents / Q000494:pharmacology; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D011853:Radiobiology; D016634:Radiosurgery / Q000379:methods*; D018874:Spheroids, Cellular / Q000187:drug effects / Q000528:radiation effects; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000528:radiation effects
chemical_list: D011837:Radiation-Protective Agents; D011838:Radiation-Sensitizing Agents; D010431:Pentoxifylline