Support is requested to define the protein map of malignant human B lymphocytes at different stages of maturation and to identify differentiation-related proteins. Total cellular proteins from twenty leukemia and lymphoma cell lines will be analyzed, using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), representing seven different B cell maturation stages plus normal peripheral blood lymphocytes. Cells will be cultured in the presence or absence of two Protein Kinase C (PKC) activators with potential B cell differentiating activity; 12-0 tetradecanoylphorbol 13 acetate (TPA) and the Bryostatins for 120 hrs. 2D- PAGE will be conducted at two time points; 72 and 129 hrs. Elsie 4 Computer System will be used to analyze, measure and identify individual protein spots in different gels according to their isoelectric point, molecular masses, and integrated densities. This will enable us to detect qualitative and quantitative changes in cellular proteins induced by either agent in comparison with untreated (control cells) in each cell line. In order to related specific protein changes to differentiation, cells will also be analyzed by immunophenotypic markers at corresponding time points using stage-restricted B cell monoclonal antibodies and flow cytometry (FCM). DNA will also be stained with propidium iodide for cell cycle analysis by FCM to determine the growth-inhibitory capacity of each agent as a requirement for differentiation. The identification of differentiation-related proteins is an important step toward the uncovering of genetic mechanisms through which differentiation agents exert their effects.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29CA050715-02
Application #
3459623
Study Section
Experimental Therapeutics Subcommittee 1 (ET)
Project Start
1990-07-01
Project End
1995-04-30
Budget Start
1991-05-01
Budget End
1992-04-30
Support Year
2
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Wayne State University
Department
Type
Schools of Medicine
DUNS #
City
Detroit
State
MI
Country
United States
Zip Code
48202
Mohammad, R M; Maki, A; Pettit, G R et al. (1996) Bryostatin 1 induces ubiquitin COOH-terminal hydrolase in acute lymphoblastic leukemia cells. Enzyme Protein 49:262-72
al-Katib, A M; Mohammad, R M; Maki, A et al. (1995) Induced expression of a ubiquitin COOH-terminal hydrolase in acute lymphoblastic leukemia. Cell Growth Differ 6:211-7
Maki, A; Diwakaran, H; Redman, B et al. (1995) The bcl-2 and p53 oncoproteins can be modulated by bryostatin 1 and dolastatins in human diffuse large cell lymphoma. Anticancer Drugs 6:392-7
Mohammad, R M; al-Katib, A; Pettit, G R et al. (1994) Successful treatment of human Waldenstrom's macroglobulinemia with combination biological and chemotherapy agents. Cancer Res 54:165-8
Mohammad, R M; Hamdan, M Y; al-Katib, A (1994) Induced expression of alpha-enolase in differentiated diffuse large cell lymphoma. Enzyme Protein 48:37-44
Mohammad, R M; Maki, A; Vistisen, K et al. (1994) Protein studies of human non-Hodgkin's B-lymphoma: appraisal by two-dimensional gel electrophoresis. Electrophoresis 15:1566-72
Mohammad, R M; Vistisen, K; al-Katib, A (1994) Protein study of T and B acute lymphoblastic leukemia cell lines. Electrophoresis 15:1218-24
al-Katib, A; Mohammad, R M; Dan, M et al. (1993) Bryostatin 1-induced hairy cell features on chronic lymphocytic leukemia cells in vitro. Exp Hematol 21:61-5
Mohammad, R M; al-Katib, A; Pettit, G R et al. (1993) Differential effects of bryostatin 1 on human non-Hodgkin's B-lymphoma cell lines. Leuk Res 17:1-8
al-Katib, A; Mohammad, R M; Khan, K et al. (1993) Bryostatin 1-induced modulation of the acute lymphoblastic leukemia cell line Reh. J Immunother Emphasis Tumor Immunol 14:33-42

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