Role of sulphated polysaccharides from in Cyclosporine A-induced oxidative liver injury in rats-1

and Mg-ATPase: μmole of inorganic phosphorous liberated/min/mg protein. Group I – Control; Group II – CsA; Group III – Sulphated polysaccharides; Group IV – CsA+sulphated polysaccharides Comparisons are made between: a-Group I and Group II, III, IV; b-Group II and Group IV. The symbols (***), (**) and (*) represent statistical significance at < 0.001, < 0.01 and < 0.05, respectively.<p><b>Copyright information:</b></p><p>Taken from "Role of sulphated polysaccharides from in Cyclosporine A-induced oxidative liver injury in rats"</p><p>BMC Pharmacology 2008;8():4-4.</p><p>Published online 20 Feb 2008</p><p>PMCID:PMC2291455.</p><p></p

Histopathological findings in the liver tissue of CsA-induced and treated groups

Control and drug control groups show normal liver architecture (Figure 3a and 3c). Liver sections treated with CsA produces marked changes like inflammation around portal triad (Triaditis) with patchy microvesicular fatty degeneration (Figure 3b). Sulphated polysaccharides treated rats (Figure 3d) show considerable reduction in the pathological changes compared to CsA-induced animals.<p><b>Copyright information:</b></p><p>Taken from "Role of sulphated polysaccharides from in Cyclosporine A-induced oxidative liver injury in rats"</p><p>http://www.biomedcentral.com/1471-2210/8/4</p><p>BMC Pharmacology 2008;8():4-4.</p><p>Published online 20 Feb 2008</p><p>PMCID:PMC2291455.</p><p></p

Effect of CsA and sulphated polysaccharides on hepatic oxidants production

Values are expressed as mean ± S.D. for six animals in each group. Group I – Control; Group II – CsA; Group III – Sulphated polysaccharides; Group IV – CsA+sulphated polysaccharides. Comparisons are made between: a-Group I and Groups II, III, IV; b-Group II and Group IV. The symbols (***) and (**) represent statistical significance at < 0.001 and < 0.01, respectively.<p><b>Copyright information:</b></p><p>Taken from "Role of sulphated polysaccharides from in Cyclosporine A-induced oxidative liver injury in rats"</p><p>http://www.biomedcentral.com/1471-2210/8/4</p><p>BMC Pharmacology 2008;8():4-4.</p><p>Published online 20 Feb 2008</p><p>PMCID:PMC2291455.</p><p></p

ING1 and 5-Azacytidine Act Synergistically to Block Breast Cancer Cell Growth

<div><h3>Background</h3><p>Inhibitor of Growth (ING) proteins are epigenetic “readers” that recognize trimethylated lysine 4 of histone H3 (H3K4Me3) and target histone acetyl transferase (HAT) and histone deacetylase (HDAC) complexes to chromatin.</p> <h3>Methods and Principal Findings</h3><p>Here we asked whether dysregulating two epigenetic pathways with chemical inhibitors showed synergistic effects on breast cancer cell line killing. We also tested whether ING1 could synergize better with chemotherapeutics that target the same epigenetic mechanism such as the HDAC inhibitor LBH589 (Panobinostat) or a different epigenetic mechanism such as 5-azacytidine (5azaC), which inhibits DNA methyl transferases. Simultaneous treatment of breast cancer cell lines with LBH589 and 5azaC did not show significant synergy in killing cells. However, combination treatment of ING1 with either LBH589 or 5azaC did show synergy. The combination of ING1b with 5azaC, which targets two distinct epigenetic mechanisms, was more effective at lower doses and enhanced apoptosis as determined by Annexin V staining and cleavage of caspase 3 and poly-ADP-ribose polymerase (PARP). ING1b plus 5azaC also acted synergistically to increase γH2AX staining indicating significant levels of DNA damage were induced. Adenoviral delivery of ING1b with 5azaC also inhibited cancer cell growth in a murine xenograft model and led to tumor regression when viral concentration was optimized <em>in vivo</em>.</p> <h3>Conclusions</h3><p>These data show that targeting distinct epigenetic pathways can be more effective in blocking cancer cell line growth than targeting the same pathway with multiple agents, and that using viral delivery of epigenetic regulators can be more effective in synergizing with a chemical agent than using two chemotherapeutic agents. This study also indicates that the ING1 epigenetic regulator may have additional activities in the cell when expressed at high levels.</p> </div

Combination Indices of ING1b with epigenetic chemotherapeutics.

<p>MDA-MB468 cells were treated with combinations of <b>A</b>) LBH589 plus 5azaC, <b>B</b>) adenoviral vector expressing GFP plus ING1b and LBH589 or <b>C</b>) adenoviral vector expressing GFP plus ING1b plus 5azaC at various concentrations and Combination Indexes were determined using CalcuSyn software. The Isobologram analysis showed that 5azaC plus Ad-ING1b showed the highest degree of synergy in inducing cell death of the combinations tested. Data used to generate this graph represent a subset of the combinations shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043671#pone-0043671-g002" target="_blank">Figure 2</a>.</p

Apoptosis in response to 5azaC and ING1b in MDA-MB468 cells.

<p><b>A</b>) Cells were pretreated with 40 µM 5azaC for 48 hours and then infected with adenoviral constructs expressing GFP or ING1b plus GFP (30 MOI). Untreated cells and cells infected with only Ad-GFP or Ad-ING1b plus GFP served as controls. Cells were harvested 24 hours later, stained for Annexin V and the percentage of the cell population undergoing apoptosis was estimated by determining the percentage of Annexin positive cells by flow cytometry. Cells treated with a combination of 5azaC and Ad-ING1b plus GFP showed higher amount of apoptosis induced in comparison to controls. One way ANOVA and Tukey’s multiple comparison post-test were performed to calculate P values comparing treated to untreated control cells (*** indicates P<0.0001). <b>B</b>) Clear morphological changes were noted in cells 24 hours after treatment with Ad-ING1b alone, or in combination with 5azaC. The combination blocked cell growth and induced morphological changes consistent with apoptosis in the great majority (99%+) of cells examined. Infection with Ad-GFP had little effect upon cell number or shape while 5azaC inhibited cell growth but was not effective in inducing an apoptotic phenotype in the majority of cells. <b>C</b>) Cells treated with the indicated agents for 48 hours were harvested in Laemmli sample buffer, lysates were boiled and equal amounts of protein from each sample were electrophoresed and blotted with the indicated antibodies.</p