Abstract P4-02-16: High miRNA26A1 and Low EZH2 Expression Levels Are Associated with Favorable Outcome to Tamoxifen in Advanced Breast Cancer through Similar Molecular Pathways

Abstract

Background: We showed that decreased expression levels of EZH2 are associated with a favorable outcome to tamoxifen in advanced breast cancer. Furthermore, EZH2 knockdown in MCF7 cells resulted in estrogen receptor (ER) upregulation and increased sensitivity to anti-estrogens. Recently, EZH2 has been identified as a target of miRNA26A1 and miRNA101. Objective: To associate miRNA26A1 and miRNA101 expression levels with: A) EZH2 and B) molecular pathways and C) outcome to first-line tamoxifen monotherapy for advanced disease. Materials & Methods: Expression levels of miRNA26A1, miRNA101, EZH2 and references (miRNA-132 and miRNA-374) were measured using quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) in 235 ER-positive primary breast cancer specimens from patients with advanced disease. The levels of expression were related to clinicopathologic factors and disease outcome. Pathway analysis was performed in a subset of 65 ER-positive tumors with available gene expression microarray data available. Computations were performed with STATA and P-values <0.05 were considered statistically significant. Results: The miRNA26A1 levels were significantly associated with levels of ER, progesterone (PgR), HER2 and EGFR, whereas miRNA101 levels showed significant relations with PgR expression and menopausal status. The miRNA26A1 and miRNA101 levels showed an inverse relation with EZH2 mRNA levels (Spearman Rank Correlation of -0.21 and -0.15, respectively, P<0.05). As continuous variable in univariate analysis, miRNA26A1 (Hazard Ratio (HR) =0.13, 95 % CI: 0.06-0.28) correlated with Time to Progression (TTP), while miRNA101 did not (HR=0.87, 95% CI: 0.70-1.07). In multivariate analysis including traditional predictive factors, the third with highest miRNA26A1 levels (HR=0.49, 95% CI: 0.34-0.72) alone, or combined with the third with lowest EZH2 levels (HR=0.56, 95% CI: 0.35-0.88) were associated with a favorable TTP independently of traditional factors. Pathway analyses identified 2 overlapping, cell cycle, related pathways with two genes (CCNE1and CDC2) differentially expressed (P<0.05) between tumors with high and low levels of miRNA26A1 and EZH2, respectively. Levels of CCNE1 and CDC2 were tested in the 235 tumors and showed as continuous variables also a significant association with TTP (HR=1.27, 95% CI: 1.12-1.45 and HR=1.53, 95% CI: 0.129-1.81). Conclusions: The miRNA26A1 and miRNA101 levels have an inverse relation with levels of EZH2, however, only miRNA26A1 has predictive value in advanced breast cancer. Pathways comparison between miRNA26A1 and EZH2 identified 2 overlapping cell cycle related pathways and the genes CCNE1 and CDC2. Low levels of EZH2, CCNE1 and CDC2 and high levels of miRNA26A1 are associated with a favorable outcome to tamoxifen therapy. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P4-02-16.