Research Paper Volume 11, Issue 20 pp 8860—8878

Long non-coding SBF2-AS1 acting as a competing endogenous RNA to sponge microRNA-142-3p to participate in gemcitabine resistance in pancreatic cancer via upregulating TWF1

Figure 6. Analysis of binding relationship between SBF2-AS1 and miR-142-3p, and expression of TWF1 in parent cells and corresponding drug resistant cells and validation of targeting relationship between miR-142-3p and TWF1. (A) Detection of the expression of miR-142-3p in parental and drug-resistant pancreatic cancer cells by RT-qPCR. (B) Detection of the expression of miR-142-3p in different cells after intervention of SBF2-AS1 by RT-qPCR. (C) Detection of gemcitabine sensitivity in parental cells overexpressing miR-142-3p by MTT assay. (D) Bioinformatics website predicts binding sites of SBF2-AS1 and miR-142-3p. (E) Dual luciferase reporter gene assay verifies the regulatory relationship between SBF2-AS1 and miR-142-3p. (F) RNA-pull down assay to verify the binding relationship between SBF2-AS1 and miR-142-3p in pancreatic cancer cells. (G) Detection of TWF1 mRNA and protein expression in parental cells and corresponding drug-resistant cells by RT-qPCR and western blot analysis. (H) RT-qPCR and western blot analysis used to detect the expression of TWF1 in transfected cells. (I) Bioinformatics software predicts the targeting relationship between miR-142-3p and TWF1. (J) Luciferase activity determination validates the targeting relationship between miR-142-3p and TWF1. Repetitions = 3; Data was analyzed using the t test or one-way ANOVA.