Novel miR-490-3p/hnRNPA1-b/PKM2 axis mediates the Warburg effect and proliferation of colon cancer cells via the PI3K/AKT pathway

Figure 5 MiR-490-3p could target binding to heterogeneous ribonucleoprotein A1-b and inhibit the Warburg effect to attenuate colon cancer cell proliferation. A: The binding sites of miR-490-3P and heterogeneous ribonucleoprotein A1 (hnRNPA1) were predicted using TargetScan; B: The wild-type (WT) and mutant type (MUT) hnRNPA1-b 3’ untranslated region reporter plasmids were co-transfected with miR-490-3P control or mimics in HEK-293T cells; C: Quantitative real-time polymerase chain reaction analysis was performed to analyze hnRNPA1-b, PKM1 and PKM2 expression in SW620 and HCT116 cells transfected with miR-490-3P control or mimics or inhibitor; D and E: SW620 and HCT116 cells were transfected with miR-490-3P control or mimics or in combination of hnRNPA1-b overexpression plasmid for 48 h. Then glucose uptake and lactate production were evaluated. Scale bar: 100 μm; F: Cell viability of SW620 and HCT116 cells transfected with miR-490-3P control or mimic or conjugated with hnRNPA1-b overexpression plasmid for 72 h was detected by the cell counting kit-8 assay; G: 5-ethynyl-2’-deoxyuridine incorporation assays were performed using SW620 and HCT116 cells with miR-490-3P control or mimics or in combination with hnRNPA1-b overexpression plasmid. The percentage of proliferating cells (red fluorescence) to total cells (blue fluorescence) was calculated. Scale bar: 100 μm; H and I: Flow cytometry was performed to evaluate the effect of miR-490-3P control or mimics or in combination with hnRNPA1-b overexpression plasmid on the cell cycle distribution and apoptosis. ^aP < 0.001 vs miR-NC + HNRNPA1-B-WT; ^bP < 0.01 vs miR-NC + HNRNPA1-B-MUT1; ^cP < 0.01 vs miR-NC + HNRNPA1-B-MUT2; ns: P > 0.05 vs miR-NC + HNRNPA1-B-MUT3; ^dP < 0.05, ^eP < 0.01, ^fP < 0.001 vs mimics-NC; ^gP < 0.001 vs inhibitor-NC; ^hP < 0.01, ⁱP < 0.001 vs mimics, n = 3. NC: Normal control; OE-A1-b: Overexpression of hnRNPA1-b.