Journal: European Urology

Article Title: The Early Effects of Rapid Androgen Deprivation on Human Prostate Cancer

doi: 10.1016/j.eururo.2015.10.042

Figure Lengend Snippet: ESR1 is upregulated in response to degarelix treatment. (A) Dot plots to show mRNA expression levels of ESR1 in untreated and degarelix-treated prostate cancer (PCa) samples by expression array. (Bi) Using the Gene Set Enrichment Analysis (GSEA) to examine the distribution of known genes with the ESR1 binding motif, shown within their promoter regions, we found that these genes were enriched among genes that degarelix treatment upregulated when they were ranked by their statistical significance. The ESR1 binding motif analysed was described previously . (Bii) This ESR1 binding motif closely matches the validated, experimentally derived ESR1 binding motif shown here. (C) GSEA demonstrates enrichment of factors known to be involved with ESR1 signalling (from the National Cancer Institute BIOCARTA curated database) among genes differentially expressed in response to degarelix ranked by statistical significance. For this analysis, the degree to which the genes were enriched is defined by the running sum statistic called the normalised enrichment score , which was 2.036 (false discovery rate q -value of 0.027; p = 0.024). (D) Representative images of PCa samples stained by immunohistochemistry (IHC) for ESR1 in untreated and degarelix-treated patients with intense nuclear staining seen in the malignant epithelia of the treated but not the untreated samples. Scale bars = 250 μm. (E) IHC ESR1 staining was increased in treated (+) compared with untreated (−) PCa samples in malignant epithelia but not cancer-associated stroma or benign epithelia (samples in triplicate; sample sizes [ n ] indicate the number of subjects [20 or 27] times 3 minus missing or damaged samples on the human tissue microarray; staining intensity 0 [none] to 3 [strong]). Mean intensities from replicate samples for each patient were used to calculate statistical significance when comparing treated and untreated groups (Mann-Whitney test). (F) Graph shows correlation between intensity of staining by IHC for ESR1 and Ki-67. Spearman's ρ correlation coefficient of 0.338 ( p = 0.041).

Article Snippet: We used Gene Set Enrichment Analysis (GSEA) version 2.0.14 (The Broad Institute, Cambridge, MA, USA).

Techniques: Expressing, Binding Assay, Derivative Assay, Staining, Immunohistochemistry, Microarray, MANN-WHITNEY