Mcl-1 Is a Novel Target of miR-26b That Is Associated with the Apoptosis Induced by TRAIL in HCC Cells

Figure 2

Mcl-1 mRNA 3′-UTR is the direct target of miR-26b. Three independent experiments were performed. (a) A predicted binding site of miR-26b in 3′-UTR of human Mcl-1 mRNA. (b) HepG2 cells were cultured in 48-well plates and were cotransfected with the reporter plasmids (pMIR-Mcl-1, pMIR-Mcl-1-M, or empty pMIR) and RNA oligos (miR-26b mimics or NC oligo) for 24 h. Then the luciferase activity was determined with Dual-Luciferase Reporter System. The expression of the reporter containing wild type 3′-UTR of Mcl-1 was suppressed by miR-26b, but not in the mutated construct or empty plasmid. . (c) qPCR and western blot analysis for Mcl-1 levels in HepG2 cells after transfection with RNA oligos (miR-26b mimics or NC oligo) and plasmid (pEGFP-N1 or pEGFP-Mcl-1) for 24 h. The suppression of Mcl-1 by miR-26b was abolished by transfection of the cells with pEGFP-Mcl-1 without 3′-UTR. .

(a)

(b)

(c)