Antiproliferative effect of ZA in the absence or presence of different concentrations of estradiol.

<p>Antiproliferative effect of rising concentrations of ZA (0, 6.25 and 12.5μM) in the three cell lines MCF-7 (A), T47D (B) and MDA-MB-231 (C) in the absence (black bars) or the presence of 1nM (grey bars) and 10nM (white bars) estradiol (EZ4U).</p

Dose-dependent antiproliferative effect of ZA.

<p>Dose-dependent antiproliferative effect of ZA on the three cell lines in the absence (A) and the presence (B) of 1nM estradiol (EZ4U). Abbreviations: OD = optical density.</p

Pro-apoptotic effect of ZA measured by Annexin/PI staining.

<p>Pro-apoptotic effect of rising concentrations of ZA in the three cell lines MCF-7 (A), T47D (B), MDA-MB-231 (C) in the absence or the presence of 1nM estradiol measured by AnnexinV/PI staining.</p

Effect of ZA on DNA synthesis.

<p>Effect of rising concentrations of ZA on DNA synthesis in the three cell lines MCF-7 (A), T74D (B) and MDA-MB-231 (C) in the absence (blue bars) and the presence (red bars) of 1nM estradiol (BrdU).</p

Proliferative effect of estradiol.

<p>Proliferative effect of 0, 1 and 10nM estradiol on the three cell lines MCF-7 (black), T74D (grey) and MDA-MD-231 (white) in the absence of ZA (EZ4U). Abbreviations: OD = optical density.</p

Cell cycle analysis by DNA content.

<p>Dose-dependent pro-apoptotic effect of ZA in the three cell lines MCF-7 (A), T47D (B), MDA-MB-231 (C) in the absence or the presence of 1nM estradiol shown by the Sub-G1-Peak.</p

Patients characteristics of the healthy control group (wildtype and <i>BRCA1</i>) and patients tumor characteristics of the group of ovarian cancer patients (wildtype and <i>BRCA1</i>).

<p>Patients characteristics of the healthy control group (wildtype and <i>BRCA1</i>) and patients tumor characteristics of the group of ovarian cancer patients (wildtype and <i>BRCA1</i>).</p

Diagnostic markers for the detection of ovarian cancer in <i>BRCA1</i> mutation carriers

<div><p>Background</p><p>Screening for ovarian cancer (OC) in women at high risk consists of a combination of carbohydrate antigen 125 (CA125) and transvaginal ultrasound, despite their low sensitivity and specificity. This could be improved by the combination of several biomarkers, which has been shown in average risk patients but has not been investigated until now in female <i>BRCA</i> mutation carriers.</p><p>Methods</p><p>Using a multiplex, bead-based, immunoassay system, we analyzed the concentrations of leptin, prolactin, osteopontin, insulin-like growth factor II, macrophage inhibitory factor, CA125 and human epididymis antigen 4 in 26 healthy wild type women, 26 healthy <i>BRCA1</i> mutation carriers, 28 wildtype OC patients and 26 OC patients with <i>BRCA1</i> mutation.</p><p>Results</p><p>Using the ROC analysis, we found a high overall sensitivity of 94.3% in differentiating healthy controls from OC patients with comparable results in the wildtype subgroup (sensitivity 92.8%, AUC = 0.988; p = 5.2e-14) as well as in <i>BRCA1</i> mutation carriers (sensitivity 95.2%, AUC = 0.978; p = 1.7e-15) at an overall specificity of 92.3%.</p><p>The used algorithm also allowed to identify healthy <i>BRCA1</i> mutation carriers when compared to healthy wildtype women (sensitivity 88.4%, specificity 80.7%, AUC = 0.895; p = 6e-08), while this was less pronounced in patients with OC (sensitivity 66.7%, specificity 67.8%, AUC = 0.724; p = 0.00065).</p><p>Conclusion</p><p>We have developed an algorithm, which can differentiate between healthy women and OC patients and have for the first time shown, that such an algorithm can also be used in <i>BRCA</i> mutation carriers. To clarify a suggested benefit to the existing early detection program, large prospective trials with mainly early stage OC cases are warranted.</p></div

Sensitivity and specificity.

<p>Sensitivity and specificity of the combination of five biomarkers (CA125, MIF, Leptin, HE4, OPN) in differentiating between healthy women and OC patients (A), healthy wildtype women and wildtype OC patients (B), healthy <i>BRCA1</i> mutation carriers and <i>BRCA1</i> mutation carriers with OC (C), healthy wildtype women and healthy <i>BRCA1</i> mutation carriers (D) and wildtype OC patients and <i>BRCA1</i> mutation carriers with OC (E).</p

Correlations of the levels of the investigated biomarkers with age in the four study groups.

<p>Correlations of the levels of the investigated biomarkers with age in the four study groups.</p