Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting sphingosine kinase 2 (SphK2) by ABC294640 inhibits colorectal cancer cell growth in vitro and in vivo

doi: 10.1186/s13046-015-0205-y

Figure Lengend Snippet: The effect of ABC294640 on SphK activity, S1P or ceramide content in CRC cells. The relative SphK activity ( a and b ), S1P content ( c ) or ceramide level ( d ) (vs. Control group) with indicated ABC294640 treatment were presented. The effect of S1P (5 μM), C6 ceramide (C6, 5 μM) or SKI-II (5 μM) on ABC294640 (1 μM)-induced HT-29 cell growth inhibition and cell death were tested by MTT assay ( e ) and LDH release assay ( f ), respectively. Mean values ± SD of three independent experiments were reported. Statistical analysis was performed comparing treatment groups with vehicle control group (“C”). *P < 0.05. # P < 0.05 vs. ABC294640 only group ( e and f )

Article Snippet: Cell permeable short-chain C6 ceramide was obtained from Avanti Polar Lipids, Inc. (Alabaster, AL).

Techniques: Activity Assay, Control, Inhibition, MTT Assay, Lactate Dehydrogenase Assay

Journal: British Journal of Pharmacology

Article Title: The ceramide kinase inhibitor NVP-231 inhibits breast and lung cancer cell proliferation by inducing M phase arrest and subsequent cell death

doi: 10.1111/bph.12886

Figure Lengend Snippet: Effect of NVP-231 and NVP-995 on CerK activity in transfected MCF-7 cells. (A) MCF-7 cells that were either left untransfected (white column, −) or transfected with a cDNA of human CerK (hCerK, +) were treated for 2 h with the indicated concentrations of NVP-231 (grey columns) or NVP-995 (black column) in the presence of NBD-C6-ceramide as described in the Methods section. Cells were harvested and taken for lipid extraction. Lipids were separated by TLC and analysed on a fluorescence imaging system. The density of spots corresponding to phosphorylated NBD-ceramide was measured and results expressed as % of CerK activity compared with Wt control cells. Data are means ± SD (n = 4). (B and C) MCF-7 cells were treated for 24 h with the indicated concentrations of NVP-231. Lipids were then extracted and taken for LC-MS/MS to quantify C16-C1P (B) and the various ceramide subspecies (C). Results are expressed as pmol lipids per 106 cells and are means ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 considered statistically significant when compared with the control samples; ###P < 0.001 statistically significant when compared with the hCerK overexpressed untreated samples.

Article Snippet: C 6 -ceramide was from Enzo Life Sciences AG (Lausen, Switzerland).

Techniques: Activity Assay, Transfection, Fluorescence, Imaging, Liquid Chromatography with Mass Spectroscopy

Journal: British Journal of Pharmacology

Article Title: The ceramide kinase inhibitor NVP-231 inhibits breast and lung cancer cell proliferation by inducing M phase arrest and subsequent cell death

doi: 10.1111/bph.12886

Figure Lengend Snippet: Tables of Links

Article Snippet: C 6 -ceramide was from Enzo Life Sciences AG (Lausen, Switzerland).

Techniques:

Journal: mBio

Article Title: Identification of a New Class of Antifungals Targeting the Synthesis of Fungal Sphingolipids

doi: 10.1128/mBio.00647-15

Figure Lengend Snippet: Measurements of sphingolipids upon treatment with BHBM. (a) Thin-layer chromatography analysis of sphingolipids (see diagram) isolated from untreated or BHBM-treated C. neoformans cells after in vivo labeling with [ 3 H]palmitate ( 3 H-Palm). (b to h) Lipid analysis by LC-MS. (b) C18 hydroxy (Δ8) 9 methyl-glucosylceramide (GlcCer); (c) C18 dihydroceramide (C18 dhCer); (d) sphingosine and dihydrosphingosine (SPHs); (e) sphingosine-1-phosphate and dihydrosphingosine-1-phosphate (SPH-1-P); (f) C18 hydroxyceramide (C18 OH-Cer); (g) C18 hydroxy Δ8-ceramide (C18 OH-Δ8-Cer); and (h) C18 hydroxy Δ8, 9 methyl-ceramide (C18 OH-Δ8, 9Me-Cer). Values that are significantly different are indicated as follows: *, P < 0.05 comparing treated to untreated (no drug). Statistical analysis was performed using analysis of variance (ANOVA) test. Statistical significance is accepted at a P value of <0.05. Data were compiled from three independent experiments.

Article Snippet: Fluorescence intensity of yeasts labeled with NBD C6-ceramide were compared using a FACScan flow cytometer (BD Biosciences, Franklin Lakes, NJ).

Techniques: Thin Layer Chromatography, Isolation, In Vivo, Labeling, Liquid Chromatography with Mass Spectroscopy

Journal: mBio

Article Title: Identification of a New Class of Antifungals Targeting the Synthesis of Fungal Sphingolipids

doi: 10.1128/mBio.00647-15

Figure Lengend Snippet: Effect of BHBM on Golgi morphology in C. neoformans and yeast-to-hyphal differentiation in C. albicans . (A) Control or BHBM-treated cells were stained with NBD C6-ceramide (Golgi and Golgi-derived compartments, green fluorescence). DIC, differential interference contrast; NBD-Cer, NBD 6-ceramide. Bars, 5 µm. (B) FACS analysis shows NBD C6-ceramide accumulation in C. neoformans . The gray histogram corresponds to C. neoformans yeast in the absence of NBD C6-ceramide. CT, control (no BHBM treatment). (C) Percentage of yeast and hyphal forms under control conditions and after BHBM treatment. Statistical analysis was performed using analysis of variance (ANOVA) test. Statistical significance is accepted at a P value of <0.05. The images in panels A and B are representative of three separate experiments. The data in panel C were compiled from three independent experiments.

Article Snippet: Fluorescence intensity of yeasts labeled with NBD C6-ceramide were compared using a FACScan flow cytometer (BD Biosciences, Franklin Lakes, NJ).

Techniques: Staining, Derivative Assay, Fluorescence