Fig. 1.
Fig. 1. Monoglycerides induce cell death in various leukemic cell lines and in primary leukemic B-CLL cells. / (A) Several human cancer cell lines corresponding to either T-cell leukemia (Jurkat and DU-528), promyelocytic (HL-60) or myelocytic (KU-812) leukemia, erythroleukemia (K-562), promonocytic (U-937) or monocytic (THP-1) leukemia, B-cell lymphoma (Raji), mammary adenocarcinoma (MCF-7), prostate adenocarcinoma (PC-3), or endometrial adenocarcinoma (HEC-1A) were incubated with the vehicle alone (1% ethanol; Ctrl; gray bars), 60 μM (hatched bars), 80 μM (white bars), or 100 μM (black bars) 1-C18:1 monoglyceride (MG). After 24-hour (left panel) or 48-hour (right panel) stimulation, cells were stained with MC540. Results are shown as mean percentages of MC540-positive cells ± SD obtained from triplicates. (B) The most susceptible cell lines from panel A were stimulated with the vehicle (Ctrl; gray bars), 100 μM glycerol (Glyc; white bars), or 100 μM oleic acid (OA; black bars) for 24 hours and stained with MC540. Results are shown as mean percentages of MC540-positive cells ± SD obtained from triplicates. Noteworthy is the fact that controls with the vehicle (1% ethanol) gave similar background as medium alone (not shown). (C) Purified B cells obtained from B-CLL patients or total normal PBMCs obtained from a healthy donor were incubated with the vehicle (Ctrl; gray bars), 60 μM (hatched bars), 80 μM (white bars), or 100 μM (black bars) 1-C18:1 monoglyceride (MG; top panel), 1-C16:1 monoglyceride (middle panel), or 3-C16:0 monoglyceride (bottom panel). Following a 3-hour treatment, cells were stained with annexin V. Results are shown as mean percentages of annexin V–positive cells ± SD from triplicates. Similar results were obtained for normal PBMCs from 3 different healthy individuals. (D) B-CLL cells obtained from patient no. 5 were incubated for 3 hours with the vehicle (Ctrl), 100 μM glycerol (Glyc), 80 μM oleic acid (OA), or 80 μM 1-C18:1 monoglyceride (MG). The cells were labeled with annexin V and PI. Flow cytometric analysis of forward scatter (FS) versus side scatter (SS) and annexin V versus PI profiles are shown for each condition. Each profile is a representative from triplicates. (E) B-CLL cells from patient no. 5 were incubated for 6 hours with the vehicle (Ctrl), 100 μM glycerol (Glyc), 80 μM oleic acid (OA), or 80 μM 1-C18:1 monoglyceride (MG), either in the absence (white bars) or the presence (black bars) of 50 μM zVAD-fmk and labeled by the TUNEL assay. Results are shown as mean percentages of TUNEL-positive cells ± SD from triplicates.

Monoglycerides induce cell death in various leukemic cell lines and in primary leukemic B-CLL cells.

(A) Several human cancer cell lines corresponding to either T-cell leukemia (Jurkat and DU-528), promyelocytic (HL-60) or myelocytic (KU-812) leukemia, erythroleukemia (K-562), promonocytic (U-937) or monocytic (THP-1) leukemia, B-cell lymphoma (Raji), mammary adenocarcinoma (MCF-7), prostate adenocarcinoma (PC-3), or endometrial adenocarcinoma (HEC-1A) were incubated with the vehicle alone (1% ethanol; Ctrl; gray bars), 60 μM (hatched bars), 80 μM (white bars), or 100 μM (black bars) 1-C18:1 monoglyceride (MG). After 24-hour (left panel) or 48-hour (right panel) stimulation, cells were stained with MC540. Results are shown as mean percentages of MC540-positive cells ± SD obtained from triplicates. (B) The most susceptible cell lines from panel A were stimulated with the vehicle (Ctrl; gray bars), 100 μM glycerol (Glyc; white bars), or 100 μM oleic acid (OA; black bars) for 24 hours and stained with MC540. Results are shown as mean percentages of MC540-positive cells ± SD obtained from triplicates. Noteworthy is the fact that controls with the vehicle (1% ethanol) gave similar background as medium alone (not shown). (C) Purified B cells obtained from B-CLL patients or total normal PBMCs obtained from a healthy donor were incubated with the vehicle (Ctrl; gray bars), 60 μM (hatched bars), 80 μM (white bars), or 100 μM (black bars) 1-C18:1 monoglyceride (MG; top panel), 1-C16:1 monoglyceride (middle panel), or 3-C16:0 monoglyceride (bottom panel). Following a 3-hour treatment, cells were stained with annexin V. Results are shown as mean percentages of annexin V–positive cells ± SD from triplicates. Similar results were obtained for normal PBMCs from 3 different healthy individuals. (D) B-CLL cells obtained from patient no. 5 were incubated for 3 hours with the vehicle (Ctrl), 100 μM glycerol (Glyc), 80 μM oleic acid (OA), or 80 μM 1-C18:1 monoglyceride (MG). The cells were labeled with annexin V and PI. Flow cytometric analysis of forward scatter (FS) versus side scatter (SS) and annexin V versus PI profiles are shown for each condition. Each profile is a representative from triplicates. (E) B-CLL cells from patient no. 5 were incubated for 6 hours with the vehicle (Ctrl), 100 μM glycerol (Glyc), 80 μM oleic acid (OA), or 80 μM 1-C18:1 monoglyceride (MG), either in the absence (white bars) or the presence (black bars) of 50 μM zVAD-fmk and labeled by the TUNEL assay. Results are shown as mean percentages of TUNEL-positive cells ± SD from triplicates.

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