Of the many heat shock proteins (hsp's) preferentially synthesized after a heat shock, the concentration of hsp 70 appears to correlate best with heat resistance, either permanent or transient. The long-term goal of this project is to establish the molecular basis related to the role that hsp 70 plays in modulating cellular responses to heat and drugs, and to enable clinicians to use this knowledge in a predictive manner. During the current period, emphasis is placed on three specific aims. (1) To develop practical assays using hsp 70 to predict the thermal response, development and decay of thermotolerance in various tissues. To accomplish this, we plan to investigate various techniques to quantify this protein, and to establish correlations between the intracellular concentration of hsp 70 and the survival kinetics of thermotolerance in model systems. (2) To use both microinjection and genetic manipulation to vary intracellular concentration of hsp 70, then to probe more directly the relationship between hsp 70, transient thermotolerance, and thermal resistance. In mammalian cells, microinjection of hsp 70 and antibodies against hsp 70 will be followed by survival and cellular localization studies of the microinjected cells after heat exposure. In yeast, clones will be constructed in which the concentration of hsp 70 can be modified by expressing the cloned gene from an inducible promoter gal 1 on a 2u multicopy plasmid. Transcription of gal 1 promoter can be modulated by varying ratios of glucose to galactose in the growth medium. Again, the thermal survival response will be related to concentration of hsp 70. (3) To establish the interrelationship between heat response, thermotolerance, drug resistance, and heat-drug interactions. This work will utilize human tumor cells resistant to either heat or drugs. SYSTEMS: Mammalian cell lines (CHO, human tumor lines), murine tumors (RIF-1, SCC VII/SF) and normal tissues (bone marrow, intestine). TECHNIQUES: Cell survival, gel electrophoresis, antibody production, enzyme-linked immunosorbent assay radioimmunoassay, immunoblotting, recombinant DNA and flow cytometry.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37CA031397-11
Application #
3482212
Study Section
Special Emphasis Panel (NSS)
Project Start
1982-02-01
Project End
1995-01-31
Budget Start
1992-04-01
Budget End
1993-01-31
Support Year
11
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065
Li, Gloria C; Mak, Johnson Y (2009) Re-induction of hsp70 synthesis: an assay for thermotolerance. 1988. Int J Hyperthermia 25:249-57
Li, Xiao-Ling; Shen, Shou-Rong; Wang, Sa et al. (2002) Restoration of T cell-specific V(D)J recombination in DNA-PKcs(-/-) mice by ionizing radiation: The effects on survival, development, and tumorigenesis. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 34:149-57
Wang, S; Guo, M; Ouyang, H et al. (2000) The catalytic subunit of DNA-dependent protein kinase selectively regulates p53-dependent apoptosis but not cell-cycle arrest. Proc Natl Acad Sci U S A 97:1584-8
Kim, D; Kim, S H; Li, G C (1999) Proteasome inhibitors MG132 and lactacystin hyperphosphorylate HSF1 and induce hsp70 and hsp27 expression. Biochem Biophys Res Commun 254:264-8
Hurd, Y L; Yakovleva, T; Nussenzweig, A et al. (1999) A novel neuron-specific DNA end-binding factor in the murine brain. Mol Cell Neurosci 14:213-24
Kurimasa, A; Ouyang, H; Dong, L J et al. (1999) Catalytic subunit of DNA-dependent protein kinase: impact on lymphocyte development and tumorigenesis. Proc Natl Acad Sci U S A 96:1403-8
Wachsberger, P R; Li, W H; Guo, M et al. (1999) Rejoining of DNA double-strand breaks in Ku80-deficient mouse fibroblasts. Radiat Res 151:398-407
Kim, D; Li, G C (1999) Proteasome inhibitors lactacystin and MG132 inhibit the dephosphorylation of HSF1 after heat shock and suppress thermal induction of heat shock gene expression. Biochem Biophys Res Commun 264:352-8
Li, G C; Ouyang, H; Li, X et al. (1998) Ku70: a candidate tumor suppressor gene for murine T cell lymphoma. Mol Cell 2:1-8
Burgman, P; Ouyang, H; Peterson, S et al. (1997) Heat inactivation of Ku autoantigen: possible role in hyperthermic radiosensitization. Cancer Res 57:2847-50

Showing the most recent 10 out of 64 publications