Cisplatin and doxorubicin both lead to loss of follicle health and a reduction in follicle numbers.

<p>(<b>A</b>) Cisplatin; (<b>B</b>) Doxorubicin: (<b>i</b>) Percentage of unhealthy follicles (clear); and (<b>ii</b>) total number of follicles (shaded) in each ovary. Bars denote mean+sem; n = 5 for all groups, stars denote significant differences relative to control (*p<0.05, **p<0.01, ***p<0.001).</p

Effect of cisplatin and doxorubicin on primordial follicles, and on expression of cleaved PARP.

<p>(<b>A, B</b>)<b>:</b> Analysis of TUNEL-positive cells within primordial follicles. (<b>A</b>)<b>:</b> Total number of primordial follicles, and number of primordial follicles containing TUNEL-positive cells, in ovaries treated with cisplatin or doxorubicin. (<b>B</b>)<b>:</b> TUNEL-positive primordial follicles further categorised into percentage in which the oocyte or the granulosa cells stained positive. Bars denote mean+sem; n = 4–5; stars denote significant differences relative to control (**P<0.01). (<b>C, D</b>)<b>:</b> Cleaved PARP expression in cisplatin and doxorubicin treated ovaries. Protein expression of cleaved PARP relative to β actin (loading control) in whole newborn ovaries following 24 h of (<b>C</b>)<b>:</b> cisplatin or (<b>D</b>)<b>:</b> doxorubicin treatment. Examples of Western blots are shown in Supporting Information, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070117#pone.0070117.s003" target="_blank">Figure S3</a>. Bars denote mean+sem; n = 3; stars denote significant differences relative to control (*p<0.05, **p<0.01, ***p<0.001).</p

Representative histological sections of cultured mouse ovaries.

<p>(<b>A, B</b>)<b>:</b> TUNEL-analysis to determine apoptosis in primordial follicles. (<b>A</b>)<b>:</b> ovary section labelled with TUNEL reaction (green) and counterstained with DAPI (blue). Inset top- and bottom-right magnification images are of the respective framed areas, illustrating examples of TUNEL positive oocytes (white arrowheads) and TUNEL positive granulosa cell (white arrow) within follicles identified as at the primordial stage in (B). (<b>B</b>)<b>:</b> section in (A) subsequently stained with haematoxylin and eosin. Inset sections in (A) correspond here to degenerated oocytes (black arrowheads) and degenerated surrounding granulosa cells (black arrow) within primordial follicles. Scale bar = 50 µm. (<b>C–H</b>)<b>:</b> Photomicrographs of haemotoxylin and eosin stained sections from ovaries treated with (<b>C</b>) control, (<b>D</b>) 3 µg ml⁻¹ imatinib, (<b>E</b>) 0.5 µg ml⁻¹ cisplatin, (<b>F</b>) cisplatin and imatinib co-treatment, (<b>G</b>) 0.05 µg ml⁻¹ doxorubicin and (<b>H</b>) doxorubicin and imatinib co-treatment. Scale bars represent 25 µm. Examples of a healthy primordial follicle (arrow), healthy growing follicle (black arrowhead) and unhealthy growing follicle (white arrowhead) are shown.</p

Cisplatin and doxorubicin affect different follicular cell types.

<p>Ovaries were treated with Cisplatin or Doxorubicin. All unhealthy transitional and primary follicles were further categorized as unhealthy due to: (<b>A</b>) poor oocyte health; (<b>B</b>) poor granulosa cell health; or (<b>C</b>) both. Bars denote mean+sem; n = 5 for all groups, stars denote significant differences relative to control (*p<0.05, **p<0.01, ***p<0.001).</p

Imatinib co-treatment with cisplatin, but not doxorubicin, rescues follicle health.

<p>Control, cisplatin-treated (0.5 µg ml⁻¹) and doxorubicin-treated (0.05 µg ml⁻¹) cultured ovaries were cultured in the presence or absence of imatinib. (<b>A</b>)<b>:</b> Percentage of unhealthy follicles (clear); (<b>B</b>)<b>:</b> Total number of follicles (shaded). Bars denote mean+sem; n = 7 for all groups, stars denote significant differences relative to control (**p<0.01). (<b>C</b>)<b>:</b> Pathway by which imatinib could protect the ovary against the damaging effect of cisplatin more effectively than it could against the damaging effect of doxorubicin.</p

Cisplatin and doxorubicin affect different follicle classes.

<p>Follicles were classified as morphologically unhealthy in cisplatin and doxorubicin treated ovaries according to follicle type. Effect of (<b>A</b>) Cisplatin or (<b>B</b>) Doxorubicin on the percentage of transitional and primary follicles classified as morphologically unhealthy. Bars denote mean+sem; n = 5 for all groups, stars denote significant differences relative to control (*p<0.05, **p<0.01, ***p<0.001).</p

Det er ikkje nok å berre løfta ein - alle må løftast i Yrkesfaglærarløftet

Figure S1. Western blots showing that the reduction in RAD51 levels after cisplatin treatment in melanoma cells is not seen in other cell types. Blots support the RAD51 quantification shown in Fig. 2 of the main text. Figure S2. Images showing that RAD51 repair foci are not induced by short term cisplatin treatment of melanoma cells. Figure S3. The cisplatin-induced reduction in RAD51 levels in melanoma cells is not affected by proteasome or lysosome inhibitor treatment. Figure S4. Melanoma cells show only a weak cisplatin-induced G2 arrest. Figure S5. Specificity of antibodies to translesion synthesis DNA Polymerases. Figure S6. Western blots showing increased levels of DNA Polymerase zeta in cisplatin-treated melanoma cells. Blots support the DNA Pol ζ and Pol η expression data shown in Fig. 7 of the main text. (PDF 2393 kb

Presentation_1_RFWD3 modulates response to platinum chemotherapy and promotes cancer associated phenotypes in high grade serous ovarian cancer.pptx

BackgroundDNA damage repair is frequently dysregulated in high grade serous ovarian cancer (HGSOC), which can lead to changes in chemosensitivity and other phenotypic differences in tumours. RFWD3, a key component of multiple DNA repair and maintenance pathways, was investigated to characterise its impact in HGSOC.MethodsRFWD3 expression and association with clinical features was assessed using in silico analysis in the TCGA HGSOC dataset, and in a further cohort of HGSOC tumours stained for RFWD3 using immunohistochemistry. RFWD3 expression was modulated in cell lines using siRNA and CRISPR/cas9 gene editing, and cells were characterised using cytotoxicity and proliferation assays, flow cytometry, and live cell microscopy.ResultsExpression of RFWD3 RNA and protein varied in HGSOCs. In cell lines, reduction of RFWD3 expression led to increased sensitivity to interstrand crosslinking (ICL) inducing agents mitomycin C and carboplatin. RFWD3 also demonstrated further functionality outside its role in DNA damage repair, with RFWD3 deficient cells displaying cell cycle dysregulation, reduced cellular proliferation and reduced migration. In tumours, low RFWD3 expression was associated with increased tumour mutational burden, and complete response to platinum chemotherapy.ConclusionRFWD3 expression varies in HGSOCs, which can lead to functional effects at both the cellular and tumour levels.</p

Additional file 1: Figure S1. of Enhanced response rate to pegylated liposomal doxorubicin in high grade serous ovarian carcinomas harbouring BRCA1 and BRCA2 aberrations

Sum of squares differences (SSD) between our SNV spectrum and the fresh frozen TCGA SNV spectrum at various allele frequency threshold for variant filtering. (DOCX 53 kb

Shifting incidence and survival of epithelial ovarian cancer (1995-2014): a SurvMark-2 study.

The aim of the study is to provide a comprehensive assessment of incidence and survival trends of epithelial ovarian cancer (EOC) by histological subtype across seven high income countries (Australia, Canada, Denmark, Ireland, New Zealand, Norway, and the United Kingdom). Data on invasive EOC diagnosed in women aged 15-99 years during 1995-2014 were obtained from 20 cancer registries. Age standardized incidence rates and average annual percentage change were calculated by subtype for all ages and age groups (15-64 and 65-99 years). Net survival (NS) was estimated by subtype, age group, and 5-year period using Pohar-Perme estimator. Our findings showed marked increase in serous carcinoma incidence was observed between 1995-2014 among women aged 65-99 years with average annual increase ranging between 2.2% and 5.8%. We documented a marked decrease in the incidence of adenocarcinoma 'not otherwise specified' with estimates ranging between 4.4% and 7.4% in women aged 15-64 years and between 2.0% and 3.7% among the older age group. Improved survival, combining all EOC subtypes, was observed for all ages combined over the 20-year study period in all countries with 5-year NS absolute percent change ranging between 5.0 in Canada and 12.6 in Denmark. Several factors such as changes in guidelines and advancement in diagnostic tools may potentially influence the observed shift in histological subtypes and temporal trends. Progress in clinical management and treatment over the past decades potentially plays a role in the observed improvements in EOC survival. This article is protected by copyright. All rights reserved