Notch receptor and ligands Receptor isoforms: Notch 1-4 Ligands: jagged 1, jagged 2, delta1, delta2, delta 3, delta 4 Microenviroment of bone marrow.

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Presentation transcript:

Notch receptor and ligands Receptor isoforms: Notch 1-4 Ligands: jagged 1, jagged 2, delta1, delta2, delta 3, delta 4 Microenviroment of bone marrow stroma

Materials and methods Cell culture: Myeloma cell line: RPMI-8226, OPM-2, LP-1, NCI-H929, U266, Hodgkin cell ine:KM-H2 Burkitt lymphoma cell line: BJAB HtTA-jag10 cell (under tetracycline control) Isolation and preparation of CD19+ B cells and CD19 low CD 38 high plasma cells Enrichment of primary multiple myeloma cells Immunohistochemistry Immunoblotting RT-PCR analysis Proliferation assay

Materials and methods Cell cultures, vectors and drugs Gene transfer Interaction between myeloma cells and stroma; detection of apoptosis Cell proliferation and cell cycle distribution Elctromobility shift assay Luciferase assay Quantitative PCR of the HES-1 gene Western blotting

Results Notch-1 activated in hematologic malignant cells following contact with bone marrow stroma Activation of the Notch-1 pathway protects myeloma cells from drug-induced apoptosis Activation of Notch-1 causes up regulation of p21 and growth inhibition of myeloma cells Overexpression of Notch-1 restored drug-resistant phenotype of Notch-1 deficient myeloma cells