The target compounds were also evaluated using the MTT assay to determine their cytotoxic activity against human non-small cell lung cancer cells (PC9, PC9GR and H460) and mouse leukemic cells (Ba/F3 WT and Ba/F3T 3151).
However CDDO-Im was 10-times more active than CDDO against mouse leukemic cells, and 20-times more active against human CLL cells compared to CDDO.
CD11c+ cells isolated from mouse lung, BMDCs, monocyte-derived (MD DCs or RAW 264.7 cells (mouse leukemic monocyte/macrophage cell line) (ATCC, Manassas, VA) were treated with indicated amount of nCB for 1 or 2 days, were washed and placed in co-culture assays with or without T cells (at 1 10 ratio).
Like the original TEL-JAK2 transgenic mice [21], diseased chimeric mice showed leukemic cell accumulation in the bone marrow, spleen, lungs, and liver (data not shown).
However, while CDDO seems to reduce the numbers of malignant B cells and normal T cells to similar extents, CDDO-Im seems to be comparatively more toxic to mouse leukemic B cells than to normal T cells.
To investigate whether this point mutation changes IRF-5 subcellular localization, wild type and point-mutated form of IRF-5 were linked to GFP, stably tansfected into HEK293 cells and RAW264.7 cells (mouse leukemic monocyte macrophage cell line), and examined for leptomycin B (LMB -induced changes in subceLMB -inducedization.
Taken together, these results demonstrate the presence of endogenous Lck kinase in the mitochondria of both human and mouse leukemic T-cells.
The mouse leukemic T-cell line LSTRA overexpresses Lck kinase and mimics the aggressive form of human large granular lymphocytic leukemia [ 42].
Mouse leukemic monocyte macrophage cell line RAW 264.7 cell line was purchased from ATCC and maintained in DMEM medium complemented with 10% fetal bovine serum and 1% antibiotics at 37°C and 5% CO2.
