Fig. 4: Circ-0075305 targets and inhibits miR-708-5p activity.

a Multiple databases predicted miRNAs with binding sites for Circ-0075305 and screened for miR-708-5p in Circ-0075305 GC cell lines with stable expression and silencing by qRT-PCR. b The co-location of Circ-0075305 (green) and miR-708-5p (red) in the cytoplasm of GC cells was verified by FISH. Cell nucleuses were counterstained with DAPI (blue). c Dual-luciferase assay was conducted to confirm the structural and functional binding relationship between Circ-0075305 and miR-708-5p. d Changes in miR-708-5p in Circ-0075305 GC cell lines were detected using qRT-PCR. e Tumor formation in GC cells inoculated subcutaneously into mice. f The changes of tumor volume were recorded. g The changes of tumor weight were recorded. h The expression and distribution of Ki-67 in these tumors were detected using immunohistochemistry. i Statistical analysis of Ki-67 positive cells in tumor tissues. j Expression levels of miR-708-5p in GC tissues and adjacent normal tissues were quantified using qRT-PCR. k The association between Circ-0075305 and miR-708-5p expression levels in GC tissues was investigated using qRT-PCR. l Analysis of the impact of miR-708-5p expression on the OS rate in postoperative chemotherapy patients with GC. Data are represented as the mean ± SD and analyzed by Student’s t-test or one-way analysis of variance (ANOVA). NS, no significance, *P < 0.05, ***P < 0.001 for groups connected by horizontal lines. P-values < 0.05 were considered statistically significant. c, d, i: n = 3 per group; f, g: n = 5 per group.