Figure 4
From: EGFR conjunct FSCN1 as a Novel Therapeutic Strategy in Triple-Negative Breast Cancer
EGF regulates FSCN1 expression and activates MAP kinase signaling in TNBC. (A,B) Expression levels of FSCN1 mRNA and protein in the MDA-MB-468 (A) and MDA-MB-231 (B) cell lines treated with EGF/Gefitinib. Upper, Real-time PCR validated the expression levels of FSCN1 mRNA in the MDA-MB-468 and MDA-MB-231 cell lines. FSCN1 expression is normalized to GAPDH. The data are presented as the mean ± S.E.M. from three independent experiments, each performed in triplicate. *P < 0.01. Under, Western blot was used to analyze the expression levels of FSCN1. Total cellular protein was isolated and subjected to western blot analysis for FSCN1 expression. GAPDH was used as loading control. Bottom, The quantitative result of western blot is shown. (C,D) Transwell assays. Cells were grown and transfected with FSCN1 siRNA or a control siRNA for 2 days. Serum deprived cells were sebsequently treated with 20 ng/mL EGF or with a Vehicle for 4 h. Cells were subsequently subjected to assays. *P < 0.01. (E,F). Expression levels of FSCN1 mRNA and protein in the MDA-MB-468 (E) and MDA-MB-231 (F) cell lines treated with EGF/U0126. Upper, Real-time PCR validated the expression levels of FSCN1 mRNA in the MDA-MB-468 and MDA-MB-231 cell lines. FSCN1 expression is normalized to GAPDH. The data are presented as the mean ± S.E.M. from three independent experiments, each performed in triplicate. *P < 0.01. Under, Western blot was used to analyze the expression levels of FSCN1. Total cellular protein was isolated and subjected to western blot analysis for FSCN1 expression. GAPDH was used as loading control. Bottom, The quantitative result of western blot is shown. (G,H) Transwell assays. Cells were grown and transfected with FSCN1 siRNA or control siRNA for 2 days and treated with 100 ng/mL Gefitinib or vehicle for 45 min. Cells were subsequently subjected to assays. *P < 0.01. I. Correlation analysis of the TCGA Breast Invasive carcinoma database (TCGA, Provisional 2012) using cBioPortal show the association between MAPK1/EGFR and FSCN1 mRNA levels, in addition to ESR1 (ER)/PGR (PR)/ERBB2 (Her2)/MAPK1 and EGFR mRNA levels. P < 0.0001.
