Early Effects of PRS-Irradiation for 9L Gliosarcoma: Characterization of Interphase Cell Death

We characterized the interphase cell death of 9L gliosarcoma after high-dose-rate, low-energy photon irradiation using the Photon Radiosurgery System (PRS), a novel device for interstitial radiotherapy. Within 24 hours after irradiation with a dose of 18 Gray, 22.0 % of cells underwent metabolic cell death, whereas dead cells in controls stayed less than 5.0 % (p < 0.005). In the majority of sensitive cells, loss of membrane integrity preceded the lethal morphological changes. The response was dose-dependent over the range of 9 - 18 Gray, but saturation was obtained over 18 Gray. On the other hand, a significant (p < 0.01) increase in the number of TUNEL-positive cells with apoptotic morphology was detected 6 - 24 hours after irradiation, but the fraction remained 1.9 - 2.1 % of the population and was independent of the doses between 9 and 25 Gray. Apoptotic cells were rarely observed in the control cells (0.3 - 0.6 %). Our data indicate that single high-dose irradiation induces both necrotic and apoptotic interphase cell death in 9L gliosarcoma, but rapid cell death mostly occurs through the non-apoptotic pathway.

Key words:

9L Gliosarcoma - Radiosurgery - Photon Radiosurgery System (PRS) - Necrosis - Apoptosis - Interphase Cell Death

References

• 1 Roper P R, Drewinko B. Comparison of in vitro methods to determine drug-induced cell lethality.  Cancer Res. 1976;  36 2182-2188
• 2 Dewey W C, Ling C C, Myen R E. Radiation-induced apoptosis: relevance to radiotherapy.  Int J Radiat Oncol Biol Phys. 1995;  33 781-796
• 3 Radford I R. The level of induced DNA double-strand breakage correlates with cell killing after X-irradiation.  Int J Radiat Biol. 1986;  48 45-54
• 4 Fowler J F. The linear-quadratic formula and progress in fractionated radiotherapy.  Br J Radiol. 1989;  62 679-694
• 5 Sheline G E, Wara W M, Smith V. Therapeutic irradiation and brain injury.  Int J Radiat Oncol Biol Phys. 1980;  6 1215-1228
• 6 van der Kogel A J. Central nervous system radiation injury in small animal models. In: Gutin PH, Leibel SA, Sheline GE (eds.). Radiation injury to the nervous system New York: Raven Press 1991: 91-111
• 7 Kreth F W, Faist M, Warnke P C, Rossner R, Volk B, Ostertag C B. Interstitial radiosurgery of low-grade gliomas.  J Neurosurg. 1995;  82 418-429
• 8 Kondziolka D, Flickinger J C, Bissonette D J, Bozik M, Lunsford L D. Survival benefit of stereotactic radiosurgery for patients with malignant glial neoplasms.  Neurosurgery. 1997;  41 776-783
• 9 McDermott M W, Sneed P K, Gutin P H. Interstitial brachytherapy for malignant brain tumors.  Semin Surg Oncol. 1998;  14 79-87
• 10 Shrieve D C, Alexander E 3rd, Black P M, Wen P Y, Fine H A, Kooy H M, Loeffler J S. Treatment of patients with primary glioblastoma multiforme with standard postoperative radiotherapy and radiosurgical boost: prognostic factors and long-term outcome.  J Neurosurg. 1999;  90 72-77
• 11 Shenouda G, Souhami L, Podgorsak E B, Bahary J P, Villemure J G, Caron J L, Mohr G. Radiosurgery and accelerated radiotherapy for patients with glioblastoma.  Can J Neurol Sci. 1997;  24 110-115
• 12 Deen D F, Hoshino T, Williams M E, Muraoka I, Knebel K D, Barker M. Development of a 9L rat brain tumor cell multicellular spheroid system and its response to 1,3-bis(2-chloroethyl)-1-nitrosourea and radiation.  J Natl Cancer Inst. 1980;  64 1373-1382
• 13 Barth R F. Rat brain tumor models in experimental neuro-oncology: the 9L, C6, T9, F98, RG2 (D74), RT-2 and CNS-1 gliomas.  J Neurooncol. 1998;  36 91-102
• 14 Weizsacker M, Deen D F, Rosenblum M L, Hoshino T, Gutin P H, Barker M. The 9L rat brain tumor: description and application of an animal model.  J Neurol. 1981;  224 183-192
• 15 Biggs D S, Thompson E S. Radiation properties of a miniature X-ray device for radiosurgery.  Br J Radiol. 1996;  69 544-547
• 16 Hakim R, Zervas N T, Hakim F, Butler W E, Beatty J, Yanch J C, Biggs P J, Gall K P, Sliski A P. Initial characterization of the dosimetry and radiobiology of a device for administrating interstitial stereotactic radiosurgery.  Neurosurgery. 1997;  40 510-517
• 17 Gavrieli Y, Sherman Y, Ben-Sasson S A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation.  J Cell Biol. 1992;  119 493-501
• 18 Ijiri K, Potten C S. Response of intestinal cells of differing topographical and hierarchical status to ten cytotoxic drugs and five sources of radiation.  Br J Cancer. 1983;  47 175-185
• 19 Shinohara C, Gobbel G T, Lamborn K R, Tada E, Fike J R. Apoptosis in the subependyma of young adult rats after single and fractionated doses of X-rays.  Cancer Res. 1997;  57 2694-2702
• 20 Majno G, Joris I. Apoptosis, oncosis, and necrosis. An overview of cell death.  Am J Pathol. 1995;  146 3-15
• 21 Kondziolka D, Lunsford L D, Pandalai S, Maitz A H, Flickinger J C. Radiobiology of radiosurgery: part II. The rat C6 glioma model.  Neurosurgery. 1992;  31 280-288
• 22 Asai A, Shibui S, Barker M, Vanderlaan M, Gray J W, Hoshino T. Cell kinetics of rat 9L brain tumors determined by double labeling with iodo- and bromodeoxyuridine.  J Neurosurg. 1990;  73 254-258
• 23 Andersen A P. Postoperative irradiation of glioblastomas. Results in a randomized series.  Acta Radiol Oncol. 1978;  17 475-484
• 24 Leibel S A, Sheline G E. Radiation therapy for neoplasms of the brain.  J Neurosurg. 1987;  66 1-22
• 25 Sheline G E. Radiation therapy of brain tumors.  Cancer. 1977;  39 873-881
• 26 Stapper N J, Stuschke M, Sak A, Stüben G. Radiation-induced apoptosis in human sarcoma and glioma cell lines.  Int J Cancer. 1995;  62 58-62
• 27 Consoli U, Li H, Huang P, Kobayashi T, Andreeff M, Levin V, Zhang W. Enhancement of apoptosis in irradiated glioblastoma cells by pentoxifylline.  Oncol Report. 1997;  4 673-678
• 28 Zhivotovsky B, Joseph B, Orrenius S. Tumor radiosensitivity and apoptosis.  Exp Cell Res. 1999;  248 10-17
• 29 Aliprantis A O, Yang R B, Mark M R, Suggett S, Devaux B, Radolf J B, Klimpel G R, Godowski P, Zychlinsky A. Cell activation and apoptosis by bacterial lipoproteins through toll-like receptor-2.  Science. 1999;  285 736-739
• 30 Finkel E. Does cancer therapy trigger cell suicide.  Science. 1999;  286 2256-2258

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