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. 2018 Nov 29;10:6537–6547. doi: 10.2147/CMAR.S185789

Figure 6.

Figure 6

miR-424 targets AKT3 and PSAT1 in HCT116 and RKO cells.

Notes: (A) The expression levels of AKT3 and PSAT1 mRNA were determined in 53 matched pairs of CRC tissues and corresponding normal colon mucosa by RT-qPCR. N stands for normal and CA for CRC samples. Data are presented as fold change. (B) Correlation between AKT3 or PSAT1 and miR-424 expression in CRC patients. (C) The predicted miR-424-binding sites and sequences in the 3′UTR of AKT3 and PSAT1 mRNAs are shown. (D) Western blotting analysis of AKT3 and PSAT1 expressions in HCT116 and RKO cells transfected with miR-424 or miR-NC at 72 hours post-transfection. GAPDH was used as a loading control. (E) Analysis of luciferase activity. PmirGLO-wild-AKT3 3′UTR, pmirGLO-mutant-AKT3 3′UTR, pmirGLO-wild-PSAT1 3′UTR, or pmirGLO-mutant-PSAT1 3′UTR was cotransfected into 293 T cells with miR-424 or miR-NC. The luciferase activity was determined. Renilla luciferase activity was used as an internal control. Data represent the average of three independent experiments. NS, not significant; **P<0.01; ***P<0.001.

Abbreviations: AKT3, AKT serine/threonine kinase 3; CRC, colorectal cancer; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; miR-NC, negative control miRNA; UTR, untranslated region.