Fig. 7: Circ-0075305 hinders the stem-like properties of GC by competitively binding with β-catenin through the RPRD1A axis.

a Wild-type C57BL6 mice developed GAC at 48 weeks after MNU induction. The proportion of RPRD1A+ cells in MNU-induced mouse tumors was significantly lower than that in normal gastric tissue (body:74.3 ± 3.8%vs.9.1 ± 1.9%, P < 0.001; Sinus:76.9 ± 5.2%vs.10.2 ± 1.7%, P < 0.001). b Immunofluorescence was used to determine whether Circ-0075305 affected the nucleation of β-catenin in GC cell lines. c The nucleocytoplasmic separation experiment was utilized to gauge the consequences of RPRD1A knockdown on TCF4 and β-catenin expression in nucleus and cytoplasm. d Statistical evaluation of results presented in c. e Western blot analysis was performed to assess alterations in the expression levels of CD44, NANOG, and SOX9 after the addition of different concentrations of the TCF4 antagonist (LF3) to HGC-27 cells. f Statistical evaluation of results presented in e. g Expression levels of β-catenin-bound RPRD1A and TCF4 in GC cell lines treated with varying concentrations of the TCF4 antagonist (LF3) were detected by CO-IP. h Immunohistochemical representation of RPRD1A in GC and normal tissue. i The box plot depicted the distributions of the immunohistochemical score of RPRD1A expression in gastric and normal tissues. The lower and upper sides of the box are the lower and upper quartiles. The whiskers are the two lines outside the box, that go from the minimum to the lower quartile and then from the upper quartile to the maximum. Each dot indicated score of individual patients. j Clinical data analysis was conducted to investigate the impact of Circ-0075305 and miR-708-5p expression on the overall survival of patients with GC who received postoperative adjuvant chemotherapy. k Examination of miR-708-5p and RPRD1A’s impact on the overall survival of GC patients receiving postoperative adjuvant chemotherapy, based on patient clinical data. l The influence of Circ-0075305 and RPRD1A expression on the overall survival of GC patient. 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 < 0.01, ***P < 0.001 for groups connected by horizontal lines. P-values < 0.05 were considered statistically significant. d, f: n = 3 per group; a: n = 5 per group.