Fig 2: Effects of RGCC and the ERK inhibitor on NP-induced cell invasion and migration. (A) Cell invasion of COLO205 and COLO205/si-RGCC cells and HCT-116 and HCT-116/ov-RGCC cells after NP (10–6 M) and/or PD98059 treatment. (B) Cell migration of COLO205 and COLO205/si-RGCC cells and HCT-116 and HCT-116/ov-RGCC cells after NP (10–6 M) and/or PD98059 treatment. ns represents p > 0.05, * represents p < 0.05, *** represents p < 0.001, **** represents p < 0.0001, n = 3.

Fig 3: Expression of RGCC in CRC tissues. (A) The relative mRNA expression of RGCC in CRC cells (SW480 and COLO205) after NP (0, 10–7, and 10–6 M) treatment by qPCR. * represents p < 0.05; *** represents p < 0.001; **** represents p < 0.0001; ns represents p > 0.05, n = 3. (B) The relative protein expression of RGCC in CRC cells (SW480 and COLO205) after NP (0, 10–7, and 10–6 M) treatment by the western blot assay. * represents p < 0.05; ** represents p < 0.01, n = 3. (C) The expression of RGCC in CRC cancer tissues in the UALCAN database. Blue represents normal tissues, and red represents tumor tissues. (D) The expression of RGCC in different stages of CRC tissues in the UALCAN database. (E) The KM analysis for RGCC in CRC patients in the UALCAN database. (F) The expressions of RGCC in CRC tumor tissues and paracancerous tissues were tested by IHC analysis. (G) The IOD analysis for IHC analysis is shown as a box figure. **** represents p < 0.0001, n = 27. (H) The correlation between the expression of RGCC and the serum level of NP in CRC patients was analyzed. n = 89. (I) The relative protein expression of RGCC in tumor tissues from the mouse xenograft model of human CRC cells after NP treatment or not was tested by western blotting, C represents that the control group was treated with physiological saline, N represents that the NP treated group was treated with NP (17 mg/(kg·day)), * represents p < 0.05, n = 4.

Fig 4: Effects of RGCC and the ERK inhibitor on the NP-induced EMT process. The expressions of the EMT-related protein (E-cadherin, N-cadherin, and vimentin) in COLO205 and COLO205/si-RGCC cells (A) and HCT-116 and HCT-116/ov-RGCC cells (B) after NP (10–6 M) and/or PD98059 treatment were tested by western blotting. The expressions of ERK1/2 and p-ERK1/2 in COLO205 and COLO205/si-RGCC cells (C) and HCT-116 and HCT-116/ov-RGCC cells (D) after NP (10–6 M) and/or PD98059 treatment were tested by western blotting. ns represents p > 0.05, * represents p < 0.05, ** represents p < 0.01, **** represents p < 0.0001, n = 3.

Fig 5: VEGFA interacted with RGC‐32, and involved in the regulation of PC. (A) Analyzed the correlation of VEGFA and RGC‐32 expression in PC tissues based on data from the GEPIA and TCGA databases. (B) Investigated the interaction between VEGFA and RGC‐32 through immunoprecipitation. (C) Analysis of RGC‐32 expression in tumor and normal tissues through GEPIA and TCGA databases. (D, E) Detection of RGC‐32 expression in tumor tissues, normal tissues, PC cell lines, and normal pancreatic cells. (F, G) Examination of RGC‐32 expression in tumor tissues at different cancer stages. (H) Survival analysis based on RGC‐32 expression in PC patients from the TCGA database. (I) Evaluation of the impact of the sh‐VEGFA on RGC‐32 expression. Significance levels are denoted as (***, p < 0.001; **, p < 0.01; *, p < 0.05).