Fig. 7
From: CircFAT1 sponges miR-375 to promote the expression of Yes-associated protein 1 in osteosarcoma cells
Knockdown of miR-375 reverses shcircFAT1-induced attenuation of cell proliferation, migration, and invasion in OS cells. a The protein expression of YAP1, c-Myc, and Birc5 in HOS and 143B cells was detected by western blot analysis. Cells were co-transfected with shcircFAT1 and miR-375 sponge or control vector. Data represent the mean ± SD (n = 3). b The mRNA expression of YAP1, c-Myc, and Birc5 in HOS and 143B cells was detected by qRT-PCR analysis. Cells were transfected with control vector and shcircFAT1 with or without miR-375 sponge. Data represent the mean ± SD (n = 3). * P < 0.05 (c) The expression of YAP1, c-Myc, and Birc5 in HOS and 143B cells was detected by immunofluorescence analysis. Cells were transfected with control vector and shcircFAT1 with or without miR-375 sponge. Data represent the mean ± SD (n = 3). Scale bars = 50 μm. d Proliferation of OS cells transfected with control vector and shcircFAT1 with or without miR-375 sponge were evaluated by the CCK-8 assay. Data represents the mean ± SD of three independent experiments. e Overexpression of circFAT1 and miR-375 induced more anoikis in osteosarcoma cells compared with circFAT1 overexpression alone. HOS and 143B stable cells were subjected to suspension culture for 48 h. Anoikis rates were determined by Annexin V-FITC/PI staining and FACS. Data represents the mean ± SD (n = 3). * P < 0.05. f miR-375 downregulation stimulated the growth of circFAT1 inhibition cells, as determined by colony formation assays (details are shown in the insets). Data represent the mean ± SD (n = 3). * P < 0.05. g Effects of circFAT1 inhibition on cell migration and invasion were eliminated by miR-375 downregulation. Migration and invasion of OS cells transfected with control vector and shcircFAT1 with or without miR-375 sponge were evaluated by the Matrigel and Transwell invasion assays, respectively. Scale bars = 50 μm. h The downregulation of circFAT1 and miR-375 on cell migration capability was evaluated by a wound healing assay in HOS and 143B cells, respectively. Data represent mean ± SD (n = 3). * P < 0.05. Scale bar, 200 μm