The overall objective of this research is to use metabolic controls to inhibit potentially lethal damage recovery (PLDR) of human tumor cells in vitro. The emphasis is on the role of extracellular PH (PHe) as a modifier of: cellular energy metabolism, rates of synthesis of macromolecules, cell growth and division, and activities of drugs which inhibit repair of radiation-damaged DNA. We have found that PLDR of A549 human lung carcinoma cells is dependent on pHe. In order to elucidate underlying mechanisms, Specific Aim 1 will investigate relationships between PLDR and changes in intracellular pH (pHi), changes in rates of biochemical reactions important for synthesis of macromolecules and production of ATP, and charges in the extent of DNA damage and the rate of its repair. Recent results with the K+/H+ ionophore nigericin suggest that such drugs may be useful for selective radiosensitization and heat sensitization of tumor cells in an acidic environment (pHe 6.5- 6.8). The mechanisms by which ionophores inhibit PLDR and increase heat toxicity will be studied as a function pHe. Optimium effects of the drugs using will be sought using metabolic controls, as well as effects of ionophores on heat radiosensitization. An important objective is to increase radiosensitivity of human tumor cells at clinically relevant radiation doses. Thus treatments which inhibit PLDR when cells are given a dose of 9 Gy will then be tested for their ability to reduce the shoulder of the dose-response curve. We have found that addition of insulin to plateau Phase A549 cells causes inhibition of PLDR.
Specific Aim 2 will continue co investigate the mechanism for this effect. Responses of human tumor cells to insulin, epidermal growth factor, transferrin and insulin-like growth factors will be optimized before mechanistic studies are undertaken. It will be of particular interest to determine limits to which alterations of cellular metabolic states can increase the radiosensitizing activities of drugs such as araA, araC, caffeine and misonidazole. The above studies will use log and plateau phase A549 cells, and occasionally CHO cells as model systems. In order to evaluate the general applicability of successful in vitro protocols, Specific Aim 3 will examine colon carcinoma cells established from human tumors ar several stages of disease progression. The health relatedness of this research is to radiation therapy and to heat treatment of cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA040516-07
Application #
3180588
Study Section
Radiation Study Section (RAD)
Project Start
1985-09-30
Project End
1994-05-31
Budget Start
1991-06-01
Budget End
1992-05-31
Support Year
7
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Case Western Reserve University
Department
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
Xue, L Y; Agarwal, M L; Varnes, M E (1995) Elevation of GRP-78 and loss of HSP-70 following photodynamic treatment of V79 cells: sensitization by nigericin. Photochem Photobiol 62:135-43
Jayanth, V R; Belfi, C A; Swick, A R et al. (1995) Insulin and insulin-like growth factor-1 (IGF-1) inhibit repair of potentially lethal radiation damage and chromosome aberrations and alter DNA repair kinetics in plateau-phase A549 cells. Radiat Res 143:165-74
Jayanth, V R; Bayne, M T; Varnes, M E (1994) Effects of extracellular and intracellular pH on repair of potentially lethal damage, chromosome aberrations and DNA double-strand breaks in irradiated plateau-phase A549 cells. Radiat Res 139:152-62
Varnes, M E; Bayne, M T; Menegay, H J et al. (1993) Effect of the K+/H+ ionophore nigericin on response of A549 cells to photodynamic therapy and tert-butylhydroperoxide. Free Radic Biol Med 15:395-405
Varnes, M E; Menegay, H J; McKenna, D S (1991) Inhibition of recovery from potentially lethal radiation damage in A549 cells by the K+/H+ ionophore nigericin. Int J Radiat Oncol Biol Phys 20:281-5
Varnes, M E; Clay, M E; Freeman, K et al. (1990) Enhancement of photodynamic cell killing (with chloroaluminum phthalocyanine) by treatment of V79 cells with the ionophore nigericin. Cancer Res 50:1620-5
Biaglow, J E; Varnes, M E; Epp, E R et al. (1989) Role of glutathione in the aerobic radiation response. Int J Radiat Oncol Biol Phys 16:1311-4
Varnes, M E; Glazier, K G; Gray, C (1989) pH-dependent effects of the ionophore nigericin on response of mammalian cells to radiation and heat treatment. Radiat Res 117:282-92
Biaglow, J E; Varnes, M E; Epp, E R et al. (1989) Role of glutathione and other thiols in cellular response to radiation and drugs. Drug Metab Rev 20:1-12
Varnes, M E (1988) Inhibition of pentose cycle of A549 cells by 6-aminonicotinamide: consequences for aerobic and hypoxic radiation response and for radiosensitizer action. NCI Monogr :199-203

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