Figure 1

Relationships between Runx2, E2 and SNAI2 in BCa. (A) Runx2 was induced by treating MCF7/Rx2^dox BCa cells with dox for 48 hours, and the effect on SNAI2 protein level was assessed by Western blot analysis of nuclear extracts. (B-C) Runx2 was either induced for 48 hours by dox treatment of C4-2B/Rx2^dox PCa cells (B) or silenced by 48-hour dox treatment of T47D/shRx2^dox BCa cells (C), and the effects on SNAI2 expression were analyzed by RT-qPCR. Western blot analysis of whole cell extracts confirms the respective dox-induced induction and silencing of Runx2. (D) Correlation between SNAI2 mRNA and Runx2 activity in a cohort of 557 BCa tumors. Runx2 activity was defined as the average normalized expression of genes that Runx2 stimulated by ≥ 2 fold in the MCF7/Rx2^dox cell culture model, except SNAI2 itself. (E) Expression of SNAI2 mRNA in the same cohort, comparing tumors with high versus low ESR1/ERα levels. Asterisk (*) indicate statistically significant difference (P < 0.05) based on unpaired t-test with Welch correction of the log2-transformation of the signal intensities. (F) Analysis of the tumors with high ESR1/ERα expression for correlation between ESR1/ERα and SNAI2. The patient cohort for D-F was compiled from the publicly available GEO datasets GSE2034, GSE2603 and GSE12276 [37–39]. BCa, breast cancer; dox, doxycycline; E2, estradiol; ESR1/ER, estrogen receptor alpha; GEO, Gene Expression Omnibus; PCa, prostate cancer; RT-qPCR; reverse transcription - quantitative polymerase chain reaction; Runx2, runt related transcription factor 2; SNAI2, snail homolog 2.