Effects of caveolin-1 on the expression of epidermal growth factor in fibroblasts co-cultured with breast cancer cells / 中国组织工程研究

ABSTRACT

BACKGROUND: Caveolin-1, as the most important functional protein in caveolae, is involoved in a variety of cell biological processes, and is closely related to the occurrence and development of breast cancer. OBJECTIVE: To explore the effect of caveolin-1 on epidermal growth factor (EGF) in fibroblasts after co-cultured with breast cancer cells.METHODS: Caveolin-1 expression in fibroblast lines ESF-1 was interfered with siRNA, and the optimal effect was determined through QRT-PCR and western blot assay. (1) The optimal silencing model of ESF-1-Caveolin-1 SiRNA-N.2 was obtained, which was co-cultured with breast cancer cells BT474 as experimental group, single-cultured ESF-1 and ESF-1-Caveolin-1 SiRNA-N.2 as controls. The expression level of EGF in ESF-1 was detected by QRT-PCR at 24 and 48 hours of culture; the expression level of EGF in the culture medium was detected by ELISA at 48 and 72 hours of culture. (2) ESF-1-Caveolin-1 SiRNA-N.2 (experimental group) and ESF-1 (control group) were respectively co-cultured with BT474, and single-cultured BT474 as blank control group. The proliferation of BT474 was detected by cell counting-kit 8 assay after 24- and 48-hour culture.RESULTS AND CONCLUSION: The mRNA expression level of EGF in the experimental group was significantly higher than that in the other two groups (P ESF-1-Caveolin-1 SiRNA-N.2 group > ESF-1 group (P < 0.05). The proliferation of BT474 cells was significantly increased after co-cultured with BT474 cells and ESF-1 siRNA Caveolin-1 cells, especially with BT474 cells (P < 0.05). Our findings suggest that Caveolin-1 siRNA can promote the expression of EGF in fibroblasts, especially co-cultured with breast cancer cells. Furthermore, caveolin-1 siRNA accelerates the proliferation of breast cancer cells after co-cultured with fibroblasts.