Fig. 1: Identification of miRNA-92a as the downstream effector of ERβ in BCa.

UMUC3 cells were transduced by lentivirus to overexpress ERβ (oeERβ) or vector control (Crtl). J82 cells were transduced with lentiviral-shERβ to knock down ERβ, or -shLuc as control for the following experiments. a Cell lysates were collected from lentivirus-transduced J82 and UMUC3 cells. Western blot analysis was performed, and proteins were detected by antibody against ERβ. b The MTT assay was used to analyze cell growth in J82 and UMUC3 cells transduced as described above to overexpress or knock down ERβ. c The Matrigel invasion assays were performed in J82 and UMUC3 cells transduced as described above. d 3D invasion assay showed that more protruding structures formed in control J82 cells than in J82 shERβ cells. e Real-time PCR screening of ERβ-regulated miRNAs related to BCa progression. We compared the PCR profile of miRNAs related to cancer cell growth and invasion in UMUC3 cells with overexpressed ERβ vs. vector (Ctrl) and J82 cells with shERβ vs. shLuc control (Ctrl). f Among the miRNAs screened, miR-92a and miR-129 expression were the most up-regulated by ERβ. We transduced lentiviral-miR-92a and lentiviral miR-129 into BCa cells, and the results showed that the increased miR-92a, not miR-129, can selectively increase BCa invasion. g We used the TCGA database to analyze the BCa sample array with miR-92a expression, and the results showed that BCa tumors have higher miR-92a expression than normal bladder tissues. In c, d, and f, data are presented as the mean ± SD. *p < 0.05