中国组织工程研究2017，Vol.21Issue(20)：3202-3207,6.DOI:10.3969/j.issn.2095-4344.2017.20.015
乳腺癌基质成纤维细胞内Caveolin-1蛋白对表皮生长因子的影响
Effects of caveolin-1 on the expression of epidermal growth factor in fibroblasts co-cultured with breast cancer cells
摘要
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 < 0.05), and the EGF expression level in the ESF-1-Caveolin-1 SiRNA-N.2 group was significantly higher than that in the ESF-1 group (P < 0.05). The protein expression level of EGF was ranked as follows: experimental group > 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.关键词
组织构建/组织工程/乳腺癌/成纤维细胞/Caveolin-1/表皮生长因子/siRNA分类
医药卫生引用本文复制引用
肖亮,王莹,杨小军,杨军平..乳腺癌基质成纤维细胞内Caveolin-1蛋白对表皮生长因子的影响[J].中国组织工程研究,2017,21(20):3202-3207,6.基金项目
江西省卫计委中医药科研课题(2015A032) （2015A032）
江西省教育厅科学技术研究项目(150882)the Chinese Medicine Research Project of the Health and Family Planning Commission of Jiangxi Province, No. 2015A032 （150882）
the Science and Technology Project of the Education Department of Jiangxi Province, No. 150882 （）
