SAHA and IFNα exerted co-operative cytotoxic effects in cancer cell lines, but not in normal non-malignant cells

<p><b>Copyright information:</b></p><p>Taken from "Enhancing the anti-angiogenic action of histone deacetylase inhibitors"</p><p>http://www.molecular-cancer.com/content/6/1/68</p><p>Molecular Cancer 2007;6():68-68.</p><p>Published online 25 Oct 2007</p><p>PMCID:PMC2173905.</p><p></p> Neuroblastoma BE(2)-C, breast cancer MCF-7, and normal non-malignant lung MRC-5 fibroblasts were treated with control, 0.5 μM SAHA and/or 500 IU/ml IFNα for 72 hours. Cell viability was examined using the Alamar blue assay, measured as optical density (OD) units of absorbance, and expressed as the absorbance of treated over control samples (ie., % viable cells). * p < 0.05, ** p < 0.01, *** p < 0.001

TSA and IFNα co-operatively suppress tumour-driven angiogenesis in neuroblastoma bearing transgenic MYCN mice

<p><b>Copyright information:</b></p><p>Taken from "Enhancing the anti-angiogenic action of histone deacetylase inhibitors"</p><p>http://www.molecular-cancer.com/content/6/1/68</p><p>Molecular Cancer 2007;6():68-68.</p><p>Published online 25 Oct 2007</p><p>PMCID:PMC2173905.</p><p></p> A. Photomicrographs of neuroblastoma tumour tissue sections from homozygous MYCN transgenic mice treated with either control, TSA, IFNα, or TSA and IFNα, which were subject to immunohistochemical studies using an anti-PECAM-1 antibody. Arrows indicate PECAM-1 positive microvessels (brown colored). B. Quantitation of the number of PECAM positive microvessels per 40× high power field in neuroblastoma tumour cross-sections. *** p < 0.001

HDACI and IFNα co-operatively inhibit endothelial cell functions, and pro-angiogenic gene expression in cancer cells under hypoxic conditions

<p><b>Copyright information:</b></p><p>Taken from "Enhancing the anti-angiogenic action of histone deacetylase inhibitors"</p><p>http://www.molecular-cancer.com/content/6/1/68</p><p>Molecular Cancer 2007;6():68-68.</p><p>Published online 25 Oct 2007</p><p>PMCID:PMC2173905.</p><p></p> A. Human umbilical vein endothelial cells (HUVECs) were treated with control (Cont), 0.1 μM TSA and/or 500 IU/ml IFNα for 18 hours. Cell viability was evaluated with the Alamar blue assay. B. HUVECs were plated in BD Biosciences Fluroblok chambers and treated with control, 0.1 μM TSA and/or 500 IU/ml IFNα for 22 hours. Cells were stained with Cell Tracker Green CMFDA, migrated through chamber filters toward the chemo-attractant VEGF, and then quantified and expressed as optical density (OD) absorbance units. C. HUVECs were plated into BD BioCoat growth factor-reduced matrigel invasion chambers and treated with control, 0.1 μM TSA and/or 500 IU/ml IFNα for 18 hours. Cells which invaded through the Matrigel were fixed, stained with a Diff Quick staining kit, photographed and then quantified. D. HUVECs were plated onto growth factor-reduced Matrigel in 24 well plates and treated with control, 0.1 μM TSA and/or 500 IU/ml IFNα for 18 hours. Vascular sprouting was quantified by counting the numbers of complete branches per branching point. E. Neuroblastoma BE(2)-C cells were treated with control, 0.02 μM TSA and/or 500 IU/ml IFNα for 72 hours under hypoxic (1% O) conditions. RNA was extracted and subjected to independent semi-competitive RT-PCR analyses using trans-intron PCR primers, together with primers for the house-keeping gene β-2 microglobulin (β2M). Representative gels for each gene at the 72 hour time point were shown, and fold induction of a target gene by treatment was calculated by ascribing the ratio between the level of expression of a target gene and that of β2M as 1.0 for control treated samples. * p < 0.05, ** p < 0.01, *** p < 0.001

Absence of p21expression correlated with sensitivity to TSA and IFNα combination therapy

<p><b>Copyright information:</b></p><p>Taken from "Enhancing the anti-angiogenic action of histone deacetylase inhibitors"</p><p>http://www.molecular-cancer.com/content/6/1/68</p><p>Molecular Cancer 2007;6():68-68.</p><p>Published online 25 Oct 2007</p><p>PMCID:PMC2173905.</p><p></p> A. MCF-7, MDA-MB-468, H460, Calu-6, DU-145, LNCaP, HT-29 and Caco-2 cells were treated with control, 0.02 μM TSA, 500 IU/ml IFNα, or TSA and IFNα for 24 hours. Whole cell protein was extracted and subjected to immunoblot with an anti- p21antibody, and, an anti-actin antibody as a loading control. B. MCF-7 cells were transfected with control scrambled or p21siRNA for 8 hours, followed by treatment with control, 0.02 μM TSA and/or 500 IU/ml IFNα for 72 hours. The effect of the siRNAs on p21gene and protein expression was analysed by semi-quantitative RT-PCR with the house-keeping gene β-2-microglobulin (β2M) as a loading control or by immunoblot, with actin as a loading control. C. Cell viability was examined by the Alamar blue assay, measured as optical density (OD) units of absorbance, and expressed as percentage of absorbance for drug-treated samples over control-treated samples (% viable cells). ** p < 0.01

TSA and IFNα exerted co-operative cytotoxic effects in cancer cell lines from a range of different tissue origins, but not in normal non-malignant cells

<p><b>Copyright information:</b></p><p>Taken from "Enhancing the anti-angiogenic action of histone deacetylase inhibitors"</p><p>http://www.molecular-cancer.com/content/6/1/68</p><p>Molecular Cancer 2007;6():68-68.</p><p>Published online 25 Oct 2007</p><p>PMCID:PMC2173905.</p><p></p> Neuroblastoma [BE(2)-C], breast (MCF-7 and MDA-MB-468), lung (H460 and Calu-6), prostate (DU-145 and LNCaP), and colon (HT-29 and Caco-2) cancer cells were treated with control (Cont), 0.02 μM TSA and/or 500 IU/ml IFNα for 72 hours. Cell viability was examined using the Alamar blue assay, measured as optical density (OD) units of absorbance, and expressed as the absorbance of treated over control samples (ie., % viable cells). ** p < 0.01, *** p < 0.001. B. MRC-5 cells were treated with control, 0.02 μM TSA and/or 500 IU/ml IFNα for 72 hours, and cell viability was assessed as above. Moreover, histone protein was extracted and subject to immunoblot analysis with anti-acetylated histone H3 antibody, after 6 hour exposure to control, TSA and/or IFNα