Validation of estrogen-responsive regulation of cell cycle and DNA replication genes suppressed by estrogen receptor beta (ERβ) overexpression by real-time quantitative polymerase chain reaction

<p><b>Copyright information:</b></p><p>Taken from "Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer"</p><p>http://breast-cancer-research.com/content/9/2/R25</p><p>Breast Cancer Research 2007;9(2):R25-R25.</p><p>Published online 10 Apr 2007</p><p>PMCID:PMC1868918.</p><p></p> , , , and were selected for further validation based on their significant correlation with ERβ transcript levels, and β actin expression was assessed as a negative control. Transcript levels in T-47Dbeta cells were measured at 30 hours following estrogen treatment (+E2) or mock treatment and under induction (-TET [+ERβ]) and non-induction (+TET) conditions. Relative fold-changes were calculated using the non-induced (+TET) samples as the reference. CDC2, cell division cycle 2; CDC6, cell division cycle 6; CKS2, cell division cycle 28 protein kinase regulatory subunit 2; DNA2L, DNA replication helicase 2-like; TET, tetracycline

Two clusters of genes showing disruption of estrogen-responsive expression profiles by estrogen receptor beta (ERβ) overexpression

<p><b>Copyright information:</b></p><p>Taken from "Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer"</p><p>http://breast-cancer-research.com/content/9/2/R25</p><p>Breast Cancer Research 2007;9(2):R25-R25.</p><p>Published online 10 Apr 2007</p><p>PMCID:PMC1868918.</p><p></p> The columns represent time points arranged in chronological order, and each row represents the expression profile of a particular gene. By convention, upregulated genes are indicated by red signals and downregulated genes are indicated by green. The magnitude of change is proportional to the brightness of the signal. (b) Second cluster of genes disrupted by ERβ overexpression

Clustering of 69 tumor samples by ERβ/ESR2, CDC2, CDC6, DNA2L, and CKS2 expression profiles is associated with clinical parameters and disease outcome

<p><b>Copyright information:</b></p><p>Taken from "Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer"</p><p>http://breast-cancer-research.com/content/9/2/R25</p><p>Breast Cancer Research 2007;9(2):R25-R25.</p><p>Published online 10 Apr 2007</p><p>PMCID:PMC1868918.</p><p></p> Hierarchical clustering of tumor samples into two groups of high (black dendrogram; cluster 1) and low (red dendrogram; cluster 2) expression clusters. Clinical parameters, including relapse and death from breast cancer within 10 years of surgery and lymph node-positive (LN+) status, are indicated with a solid bar beneath each tumor sample. Tumor grade is indicated by colored bars; green, blue, and red bars denote grades 1, 2, and 3, respectively. Gray bars denote missing data. Significant distribution of clinical parameters between the two clusters was determined by Fisher exact tests, and values were derived from the calculated distributions. Kaplan-Meier plot of 10-year censored disease-free survival (DFS) curves for cluster 1 (black) and cluster 2 (red) patients is shown with the associated values from likelihood-ratio analysis. Kaplan-Meier plot of 10-year censored disease-specific survival (DSS) curves for cluster 1 (black) and cluster 2 (red) patients is shown with the associated values from likelihood-ratio analysis. CDC2, cell division cycle 2; CDC6, cell division cycle 6; CKS2, cell division cycle 28 protein kinase regulatory subunit 2; DNA2L, DNA replication helicase 2-like; ERβ/ESR2, estrogen receptor beta

Clustering of 45 previously published ERα-positive tumor samples 40 confirms that ERβ-responsive gene set expression profile is associated with disease outcome

<p><b>Copyright information:</b></p><p>Taken from "Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer"</p><p>http://breast-cancer-research.com/content/9/2/R25</p><p>Breast Cancer Research 2007;9(2):R25-R25.</p><p>Published online 10 Apr 2007</p><p>PMCID:PMC1868918.</p><p></p> Hierarchical clustering of tumor samples into two groups of high (black dendrogram) and low (red dendrogram) ERβ expression clusters. Kaplan-Meier plot of disease-free survival (DFS) curves for high (black) and low (red) expression patients is shown with the associated values. Kaplan-Meier plot of disease-specific survival (DSS) curves for high (black) and low (red) expression patients is shown with the associated values. Due to the absence of the corresponding probes on the arrays used by Sotiriou and colleagues [40], ERβ and DNA2L transcript levels were not assessed. DNA2L, DNA replication helicase 2-like; ER, estrogen receptor