Fig. 3

miR-6794-5p directly inhibits the expression of SOCS1. A Venn diagram indicated target candidate genes of miR-6794-5p using TargetScan and miRWalk, which are miRNA target prediction sites. B, C After overexpression of the miR-6794-5p mimic in U251 (B) and A549 (C) cells, the mRNA expression of each of the candidate genes was confirmed by qRT-PCR. The values were normalized to GAPDH. D After miR-6794-5p was overexpressed in both cells, the level of SOCS1 protein was confirmed by Western blot analysis. β-actin was used for normalization. The experiment was repeated with triplicates and representative Western blotting images are shown. E, F Dual luciferase activity was examined after A549 cells were co-transfected with wild-type (WT) or mutant (Mut) vectors of the SOCS1 3’UTR in the presence or absence of the miR-6794-5p mimic, respectively. G Ago2-RNA immunoprecipitation (Ago2-IP) assay was performed in negative control (NC) or miR-6794-5p overexpressed A549 cells, and SOCS1 enrichment was confirmed by qRT-PCR. The values were normalized to GAPDH. H The mRNA expression of SOCS1 in plasma of normal and patients with lung cancer (normal, n = 23; lung cancer, n = 23) was analyzed by qRT-PCR. I Kaplan-Meier plots were used to compare survival rates between normal groups and lung adenocarcinoma patients. All data are presented as the mean ± S.D. after triplicate. *P < 0.05; **P < 0.01; ***P < 0.001. Student’s t-test