Figure 5.

Depleted LINC01234 Enhances miR-31-5p-Mediated Downregulation of MAGEA3 to Prevent HCC Progression

(A) The subcellular localization of LINC01234 in HepG2 cells identified by FISH (×400). (B) The luciferase activity of LINC01234 in HepG2 cells upon miR-31-5p mimic transfection in a dual-luciferase reporter system. (C) The binding between LINC01234 and miR-31-5p detected by RNA pull-down. (D) The binding between LINC01234 and Ago2 or DICER detected by RIP. (E) MAGEA3 expression in HepG2 cells in response to altered expression of LINC01234 and/or miR-31-5p measured using qRT-PCR. (F) Binding of miR-31-5p to MAGEA3 in HepG2 cells detected using RNA pull-down. (G–I) The viability (G), invasion (×200; H), and cisplatin-induced apoptosis (I) of HepG2 cells upon inhibition of LINC01234 and/or miR-31-5p assessed using MTT, Transwell, and flow cytometry assays. (J) IC₅₀ value of HepG2 cells upon inhibition of LINC01234 and/or miR-31-5p. (K) Protein expression of MRP2, MRP3, MDR-1, and E-cadherin in HepG2 cells upon inhibition of LINC01234 and/or miR-31-5p determined using western blot analysis. (L) ALB content in the supernatant of HepG2 cells upon inhibition of LINC01234 and/or miR-31-5p detected by ELISA. Measurement data were expressed as mean ± SD. Differences among groups at different points of time were assessed with repeated-measures ANOVA. The comparison between two groups was analyzed by independent sample t test and the comparisons among multiple groups by one-way ANOVA, followed by Tukey’s post hoc test. Each experiment was repeated three times. *p < 0.05 versus the IgG immunoprecipitates or the NC- or si-ctrl-transfected cells; #p < 0.05 versus the si-LINC01234-transfected cells.