Figure 3: BRMS1L inhibits breast cancer cell invasion and EMT via suppressing FZD10 expression.

(a) mRNA microarray (GSE61354) analysis reveals mRNA differentially expressed in MDA-MB-231 cells transfected with pcDNA3 vector carrying BRMS1L (BRMS1L) or negative control (NC) for 24 h. (b) qRT–PCR for the mRNA expression of WNT2B, WNT7A, WNT9B, WNT10B, FRAT1, FZD10, VANGL2, BMP8A, CAMK3B in mRNA microarray. ***P≪0.001 as compared with NC. Bars correspond to mean±s.d. The data are representative of three independent experiments. Student’s t-test was used for the comparison of two independent groups. (c) Western blotting for BRMS1L and FZD10 in T47D cells that were untreated (UT), mock transfected (mock) or transfected with GFP-siRNA or two BRMS1L-siRNAs. (d) Western blotting for BRMS1L and FZD10 in MDA-MB-231 cells that were transfected with pcDNA3 vector carrying BRMS1L (BRMS1L) or negative control (NC). (e) E-cadherin and vimentin protein expression levels measured by western blotting in MCF-10A cells that were untreated (UT), mock transfected (mock) or transfected with GFP-siRNA or two FZD10-siRNAs. (f,g) Invasion of cancer cells measured by Boyden chamber assays for BT-474 (f) and MDA-MB-231 cells (g) treated as indicated. Bars correspond to mean±s.d. ***P≪0.001 as compared with mock. Scale bars correspond to 50 μm. Student’s t-test was used for the comparison of two independent groups. (h) Immunohistochemical staining for FZD10 in primary breast cancer with or without regional lymph nodes (LN) metastasis and metastatic lymph nodes (LNMs). Scale bar corresponds to 50 μm. BRL, BRMS1L; E-cad, E-cadherin; Vim, Vimentin.