ERα expression and intracellular levels of glycogen in SKOV-3 and SKOV-3-R.

<p>(A) Gene expression of <i>ESR1</i> evaluated by qRT-PCR (<i>n</i> = 3). The decrease in expression in SKOV-3-R was statistically significant (<i>p</i><0.001, using independent t-test). Expression levels were normalised to <i>ACTB</i>. Error bars represent S.E.M. (B) Representative western blot showing protein expression of ERα. β-tubulin was used as a loading control. (C) Glycogen levels were evaluated using a colorimetric assay (<i>n</i> = 3). The increased level in SKOV-3-R was statistically significant (<i>p</i> = 0.027, using independent t-test). Error bars represent S.E.M.</p

Comparison of ovarian HGSC and CCC.

<p>Representative hematoxylin and eosin staining of ovarian HGSC and CCC. We hypothesise that these subtypes represents extreme forms of EOC; HGSC generally display positive expression of PGC1α, TFAM and ERα, have a higher Ki-67 index and are more responsive to initial platinum/taxane chemotherapy. By contrast, CCC generally do not express PGC1α, TFAM and ERα, have a lower Ki-67 index and are less responsive to initial platinum/taxane chemotherapy. Magnification 400x, scale bar shows 50 µm.</p

Expression of PGC1α and TFAM in EOC subtypes.

<p>(A) Representative immunohistochemical staining of VDAC, ERα, PGC1α and TFAM in HGSC and CCC. VDAC indicates presence of mitochondria. ERα, PGC1α and TFAM were all expressed in HGSC, whereas in CCC expression of the proteins was lost. Each HGSC and CCC quadruplicate is from one and the same patient (magnification: 400x, scale bar shows 500 µm). (B) Histogram representing number of cases with expression of PGC1α/TFAM across EOC subtypes.</p

Expression of Mitochondrial Regulators PGC1α and TFAM as Putative Markers of Subtype and Chemoresistance in Epithelial Ovarian Carcinoma

<div><p>Epithelial ovarian carcinoma (EOC), the major cause of gynaecological cancer death, is a heterogeneous disease classified into five subtypes. Each subtype has distinct clinical characteristics and is associated with different genetic risk factors and molecular events, but all are treated with surgery and platinum/taxane regimes. Tumour progression and chemoresistance is generally associated with major metabolic alterations, notably altered mitochondrial function(s). Here, we report for the first time that the expression of the mitochondrial regulators PGC1α and TFAM varies between EOC subtypes; furthermore, we have identified a profile in clear-cell carcinoma consisting of undetectability of PGC1α/TFAM, and low ERα/Ki-67. By contrast, high-grade serous carcinomas were characterised by a converse state of PGC1α/TFAM, ERα positivity and a high Ki-67 index. Interestingly, loss of PGC1α/TFAM and ERα was found also in a non-clear cell EOC cell line made highly resistant to platinum in vitro. Similar to clear-cell carcinomas, these resistant cells also showed accumulation of glycogen. Altogether, our data provide mechanistic insights into the chemoresistant nature of ovarian clear-cell carcinomas. Furthermore, these findings corroborate the need to take into account the diversity of EOC and to develop subtype specific treatment strategies.</p></div

PGC1α and TFAM expression in EOC SKOV-3 cells and the multiresistant subline SKOV-3-R.

<p>(A) Gene expression of <i>PPARGC1A</i> and <i>TFAM</i> in SKOV-3 and SKOV-3-R cells was evaluated by qRT-PCR (<i>n</i> = 3). The decrease in expression in SKOV-3-R was statistically significant (both <i>p</i><0.001, using independent t-test). Expression levels were normalised to <i>ACTB</i>. Error bars represent S.E.M. (B) Representative western blot showing protein expression of PGC1α and TFAM. β-tubulin was used as loading control.</p

Expression status of PGC1α and TFAM in the EOC study population.

<p>Abbreviations: EOC; epithelial ovarian carcinoma, ERα; oestrogen receptor alpha, nd; not determined, ns; not significant, PR; progesterone receptor, PGC1α; Peroxisome proliferator-activated receptor gamma co-activator 1-alpha, TFAM; mitochondrial transcription factor A.</p>a<p> Across variable; tested with Fisher's exact test for subtypes, expression of ERα and PR, and Mann-Whitney U test for distribution of age and Ki-67.</p>b<p> Within variable; tested with non-parametric Chi² test.</p><p>Expression status of PGC1α and TFAM in the EOC study population.</p

Expression status of PGC1α/TFAM in EOC.

<p>Abbreviations: EOC; epithelial ovarian carcinoma, ERα; oestrogen receptor alpha, nd; not determined, ns; not significant, PR; progesterone receptor, PGC1α; Peroxisome proliferator-activated receptor gamma co-activator 1-alpha, TFAM; mitochondrial transcription factor A.</p>a<p>Across variable; tested with Fisher's exact test for subtype, expression of ERα and PR, and Kruskal-Wallis test for distribution of age and Ki-67.</p><p>Expression status of PGC1α/TFAM in EOC.</p

Ki-67 index distribution in TFAM+ and TFAM- EOC tumours.

<p>EOC tumours with negative expression of TFAM (<i>n</i> = 18) showed significantly lower Ki-67 index compared to tumours with positive expression of TFAM (<i>n</i> = 35) (Mann-Whitney U, <i>p</i> = 0.014).</p

Animals treated with oxaliplatin demonstrated reduced cell infiltration, cartilage and bone destruction as well as less cytosolic and extracellular HMGB1

Oxaliplatin treated mice (= 8 with a mean clinical score of 1.0) and mice given vehicle alone (= 8 with a mean clinical score of 4.8) were killed on day 36 PI and intra-articular effects of oxaliplatin were evaluated by immunohistochemistry. Cell infiltration , cartilage and bone destruction were all significantly lower in oxaliplatin treated mice. Representative micrographs illustrating HMGB1 staining (in brown) in synovial tissue where an abundant cytosolic and extracellular HMGB1 staining was evident in mice treated with control vehicle as compared to the low extranuclear HMGB1 expression in oxaliplatin treated mice . Sequential sections stained with Safranin O demonstrating reduced proteoglycan content in articular cartilage in the control than in oxaliplatin treated animals . Signs of pronounced articular destructions are more evident in control treated animals, where destained cartilage layers reflected loss of matrix proteoglycans (see arrow). No cartilage destruction was detected in oxaliplatin treated mice indicated by a homogenous cartilage staining. CI, cell infiltration, JC, joint cavity, B, bone, C, cartilage. The boxes represent 25th to 75th percentiles and the lines inside the boxes stand for the median. The lines outside the boxes reflect 10th and 90th percentiles and circles indicate outliers. p = 0.05. Animals: control = 8, oxaliplatin = 8. Paws: control = 32, oxaliplatin = 32.<p><b>Copyright information:</b></p><p>Taken from "Oxaliplatin retains HMGB1 intranuclearly and ameliorates collagen type II-induced arthritis"</p><p>http://arthritis-research.com/content/10/1/R1</p><p>Arthritis Research & Therapy 2008;10(1):R1-R1.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2374449.</p><p></p

Stimulated macrophages demonstrated a nuclear retention of HMGB1 in co-cultures with oxaliplatin

RAW 264.7 macrophage-like cells were cultured without exogenous stimulus or with IFN-γ and LPS alone or with IFN-γ and LPS in the presence of 0.5 μM oxaliplatin . Immunostaining for HMGB1 (green), nucleus (blue) and the cytoskeletal protein β-actin (red) was performed in fixed cells after a culture time of 24 h. Almost all nuclei demonstrated strong HMGB1 expression in resting cells , while the nuclear HMGB1 was as expected substantially reduced in IFN-γ- and LPS-activated macrophages . Macrophages that had been activated in the same way in the presence of oxaliplatin demonstrated a strong nuclear retention of HMGB1 .<p><b>Copyright information:</b></p><p>Taken from "Oxaliplatin retains HMGB1 intranuclearly and ameliorates collagen type II-induced arthritis"</p><p>http://arthritis-research.com/content/10/1/R1</p><p>Arthritis Research & Therapy 2008;10(1):R1-R1.</p><p>Published online 7 Jan 2008</p><p>PMCID:PMC2374449.</p><p></p