Figure 5.
miR-3656 targeting of RHOF modulates the EMT process in PC. (a) Representative images showing the associations between miR-3656 expression, RHOF and EMT markers (Vimentin and E-cadherin) in 157 PC FFPE samples. (b–e) The percentage scoring of EMT marker (Vimentin and E-cadherin) expression in either miR-3656 low or high expression groups, and in either RHOF low or high expression groups from 157 PC FFPE samples. (f) Morphological changes resulting from RHOF overexpression in PANC-1 cells and RHOF-deficient BXPC-3 cells via crystal violet staining. (g-i) Examination of EMT marker (E-cadherin, N-cadherin, Vimentin and TWIST1) expression in RHOF-overexpressing PANC-1 cells and RHOF low expressing BXPC-3 cells by qPCR and western blotting assay. (j) Comparison of EMT marker (E-cadherin, N-cadherin, Vimentin and TWIST1) expression between miR-3656 single overexpressing and miR-3656/RHOF co-overexpressing PANC-1 cells. (k) Expression changes of EMT markers among si-RHOF, miR-3656 overexpressing and si-RHOF/miR-3656 overexpressing BXPC-3 cells via qPCR. (l) Western blotting assay of (j and k). GAPDH was used to normalize the qPCR results, and β-actin was used as a loading control in western blots. All n=3; bar, S.E.M., *P<0.05; **P<0.01; ***P<0.001; Student’s t-test or χ2 test