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Research on the Effects of Caveolin-1 siRNA on the Expression of Autophagosome in Fibroblasts Co-cultured with Breast Cancer Cells
Shi Xiaoyu1, Xiao Liang2, Xiong Lixia1, Meng Chuang3, Qi Guanyun3, Li Wenlin3*
1College of Basic Medicine, Nanchang University, Nanchang 330006, China;2Clinical Laboratory Centre, Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China;3Jiangxi Key Laboratory of Medical Biology, Nanchang University, Nanchang 330006, China
Abstract: In order to explore the correlation between Caveolin-1 (Cav-1) and autophagosome and its role in breast cancer cells， siRNAs were used to interfere with Cav-1 expression in fibroblast line ESF in this study. qRT-PCR and Western blot were used to determine the effect of siRNA interfering Cav-1 expression in ESF cells. Breast cancer cell lines BT474 and ESF were co-cultured by Transwell insert. The effect of Cav-1 siRNA on the expression of autophagosome in ESF cells was examined by monodansylcadaverin (MDC) staining and laser confocal microscopy. The effect of Cav-1 siRNA on the expression of microtubule-associated protein 1 light chain 3 II (LC3II) in ESF cells was examined by qRT-PCR and Western blot. The proliferation and viability of BT474 cells were measured by CCK-8 assay. The results showed that Cav-1 expression in ESFsiCav-1 (ESF cells transfected with Cav-1 siRNA) was downregulated by siRNA targeting Cav-1. The expressions of LC3II and autophagosome in ESF cells were increased by Cav-1 siRNA， and enhanced significantly in the co-culture of ESFsiCav-1 and BT474 cells. The proliferation of BT474 cells was significantly faster under the condition of co-culture of BT474 cells and ESFsiCav-1 cells. These findings suggested that Cav-1 siRNA promoted the expression of LC3II and autophagosome in fibroblasts co-cultured with breast cancer cells and Cav-1 siRNA accelerated the proliferation of breast cancer cell co-cultured with fibroblasts.
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