DataSheet_1_GBM tumors are heterogeneous in their fatty acid metabolism and modulating fatty acid metabolism sensitizes cancer cells derived from recurring GBM tumors to temozolomide.zip

Glioblastoma is a highly lethal grade of astrocytoma with very low median survival. Despite extensive efforts, there is still a lack of alternatives that might improve these prospects. We uncovered that the chemotherapeutic agent temozolomide impinges on fatty acid synthesis and desaturation in newly diagnosed glioblastoma. This response is, however, blunted in recurring glioblastoma from the same patient. Further, we describe that disrupting cellular fatty acid homeostasis in favor of accumulation of saturated fatty acids such as palmitate synergizes with temozolomide treatment. Pharmacological inhibition of SCD and/or FADS2 allows palmitate accumulation and thus greatly augments temozolomide efficacy. This effect was independent of common GBM prognostic factors and was effective against cancer cells from recurring glioblastoma. In summary, we provide evidence that intracellular accumulation of saturated fatty acids in conjunction with temozolomide based chemotherapy induces death in glioblastoma cells derived from patients.</p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-0

-16 (white dots). Sample classes are glioses, anaplastic astrocytomas and glioblastoma multiformes. (B) Glioblastoma multiforme cell lines (U87 and U251) were treated with 5-aza-dC at 1 μM (Aza.1) or 5 μM (Aza.5) alone, trichostatin A (100 ng/ml) alone, or combinations of both agents. MicroRNA expression was measured relative to let-7a and normalized to vehicle control (dimethyl sulfoxide). Error bars represent standard deviation of triplicate polymerase chain reactions from a single experimental set. Similar results were obtained in independent experiments (see Additional file ).<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-6

-16 (white dots). Sample classes are glioses, anaplastic astrocytomas and glioblastoma multiformes. (B) Glioblastoma multiforme cell lines (U87 and U251) were treated with 5-aza-dC at 1 μM (Aza.1) or 5 μM (Aza.5) alone, trichostatin A (100 ng/ml) alone, or combinations of both agents. MicroRNA expression was measured relative to let-7a and normalized to vehicle control (dimethyl sulfoxide). Error bars represent standard deviation of triplicate polymerase chain reactions from a single experimental set. Similar results were obtained in independent experiments (see Additional file ).<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-5

8 hours relative to cells transfected with 100 nM control oligonucleotide. Cyclin-dependent kinase 6 expression was determined by TaqMan relative to control genes (black bars), (gray bars) and (white bars). Values represent average +/- standard deviation of independent experiments. (B) Cyclin-dependent kinase 6 protein expression is dramatically reduced as determined by western blotting following transfection of miR-124 or miR-137. Levels of phosphorylated RB (pSer 807/811) are also markedly reduced in response to miR-124 or miR-137 transfection. (C) miR-137 sequence in relation to the pMIR-REPORT vector containing the predicted cyclin-dependent kinase 6 miR-137 binding site (wild type). Mutated bases (underlined) were also introduced into the miR-137 seed region (boxed) of the cyclin-dependent kinase 6-3'UTR (mutated). Vertical lines denote Watson-Crick base pairing. (D) Relative luminescence of U251 cells following transfection miR-137 or negative control microRNA in conjunction with wild type or mutated cyclin-dependent kinase 6 reporter constructs. *P < 0.0001.<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-2

oligonucleotide. Cells were immunostained with Tuj1 and glial fibrillary acidic protein antibodies, nuclei were counterstained with 4'-6-diamidino-2-phenylindole- and images are merged. Scale bar is 10 μm. (B) Phase contrast images of subventricular zone-neural stem cells 48 hours post-transfection by miR124 and miR137 and Tuj1 immunostaining of the same cultures 72 hours post-transfection. (C) Quantification of percentage of Tuj1+ cells, Tuj1+ cells with neuronal morphology and glial fibrillary acidic protein+ cells 72 hours after transfection with miR-124, miR-137, both miR-124 and miR-137, control oligonucleotide or transfection reagent.<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-4

CroRNA concentration) miR-124, miR-137, miR-124 and miR-137 together or negative control oligonucleotides (neg#1, neg#2) to U251 (A) and SF6969 (B) glioblastoma multiforme cells. Cells were treated with bromodeoxyuridine for 30 minutes, fixed, treated with fluorescein isothiocyanate-labeled antibromodeoxyuridine antibody and the DNA stain 7-amino-actinomycin D and subject to flow cytometry. Values represent mean ± standard deviation of replicate experiments; *P < 0.05.<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-3

Leotides and lipofectamine reagent alone. Cells were immunostained with Tuj1 and glial fibrillary acidic protein antibodies and DNA was stained with Hoechst 33258 reagent. The percentage of Tuj1- and glial fibrillary acidic protein-positive cells was quantified in each sample after transfection and staining and plotted against the total number of counted cells (= 450). (C) Quantification of Tuj1+ and glial fibrillary acidic protein+ cells in primary glioblastoma multiforme cultures 10 days after transfection of miR-124, miR-137 or control oligonucleotides. The inset shows a Tuj1+ cell with neuronal morphology from a miR-124 and/or miR-137 cotransfection. (D) Immunostaining with neuronal markers Tuj1 and microtubule-associated protein 2 10 days after miR-137 or negative control miR-transfections in glioblastoma multiforme lines maintained as neurospheres.<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-7

E-neural stem cells cultures in proliferation conditions (A)-(D), and following 1 day (E)-(H), 2 days (I)-(L), 3 days (M)-(P) and 4 days (Q)-(T) of mitogen deprivation. Phase images (A), (E), (I), (M), (Q) are shown with corresponding epifluorescent images showing 4'-6-diamidino-2-phenylindole-stained nuclei (B), (F), (J), (N), (R) and Tuj1 expression (C), (G), (K), (O), (S). Glial fibrillary acidic protein expression (D), (H), (L), (P), (T) is shown in parallel cultures. (B) Expression analysis of high-grade astrocytoma-microRNAs during a 5-day differentiation time-course of subventricular zone-neural stem cells.<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p

MiR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells-1

E-neural stem cells cultures in proliferation conditions (A)-(D), and following 1 day (E)-(H), 2 days (I)-(L), 3 days (M)-(P) and 4 days (Q)-(T) of mitogen deprivation. Phase images (A), (E), (I), (M), (Q) are shown with corresponding epifluorescent images showing 4'-6-diamidino-2-phenylindole-stained nuclei (B), (F), (J), (N), (R) and Tuj1 expression (C), (G), (K), (O), (S). Glial fibrillary acidic protein expression (D), (H), (L), (P), (T) is shown in parallel cultures. (B) Expression analysis of high-grade astrocytoma-microRNAs during a 5-day differentiation time-course of subventricular zone-neural stem cells.<p><b>Copyright information:</b></p><p>Taken from "miR-124 and miR-137 inhibit proliferation of glioblastoma multiforme cells and induce differentiation of brain tumor stem cells"</p><p>http://www.biomedcentral.com/1741-7015/6/14</p><p>BMC Medicine 2008;6():14-14.</p><p>Published online 24 Jun 2008</p><p>PMCID:PMC2443372.</p><p></p