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. 2017 May 23;8(37):61327–61337. doi: 10.18632/oncotarget.18099

Figure 5. Modulation of ROBO1in HeP3B cells directly regulated by miR-218.

Figure 5

(A) Transcription level of ROBO1 and miR-218 in cell lines were detected by qRT-PCR. The mRNA level of ROBO1 is significantly higher in HCC cells than that in QSG-7701 cells (*P<0.05), and the level of miR-218 is obviously down-regulated in HCC cells compared with QSG-7701 (*P<0.05). Results above are means of three independent experiments ± SD. (B) Scheme for the predicted miR-218 binding site in the wild type ROBO1 mRNA 3’UTR (3’UTR-WT), and in the mutant construct (3’UTR-MUT). (C) The direct interaction between miR-218 and ROBO1 was detected by Dual-luciferase reporter assay. Over-expression of miR-218 in HeP3B cells (HeP3B/miR-218) significantly decreased the luciferase signal of ROBO1/pMIR/WT compared with the negative control (HeP3B/NigmiR) (*P<0.05). And mutation of the putative miR-218-binding site abolished this suppressive effect. (D) QRT-PCR was undertaken to detect the effect of miR-218 on ROBO1 mRNA level in HeP3B cells. ROBO1 expression was significantly suppressed by over-expressing miR-218 in HeP3B cells. The results are means of three independent experiments ± SD. (*P<0.05).