Figure 2.

Altered miR-3656 expression in PC cell lines modifies sensitivity to gemcitabine. (a) MTS analysis of PANC-1 and BXPC-3 cells transfected with miR-3656 antisense (As-miR-3656), or negative control antisense (As-Con), miR-3656 mimic (Mimic-miR-3656), negative control mimic (Mimic-Con). (b and c) Representative images and statistical analysis of colony formation index among PANC-1 and BXPC-3 cells transfected with miR-3656 mimic, antagomir, or negative control. (d and e) MTS assay and the corresponding IC50 of gemcitabine measured in PANC-1 and BXPC-3 cells transfected with miR-3656 mimic, antagomir, or negative control. (f) Quantification of Annexin V/PI-positive apoptotic cells in PANC-1 and BXPC-3 cells transfected with miR-3656 mimic, antagomir or negative control after gemcitabine treatment (10 nM) via flow cytometric analysis. (g) Representative images of crystal violet staining showing the morphological changes of PANC-1 cells transfected with miR-3656 antisense or control antisense and BXPC-3 cells with miR-3656 mimic or control mimic. (h and i) EMT-related marker expression in PANC-1 cells transfected with miR-3656 antisense or control antisense, and in BXPC-3 cells with miR-3656 mimic or control mimic addition shown by western blotting and qPCR. 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., n.s, not significant, *P<0.05; **P<0.01; ***P<0.001; Student’s t-test