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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Restoration of mutant K-Ras repressed miR-199b inhibits K-Ras mutant non-small cell lung cancer progression

Fig. 4

miR-199b targets several components of the RAS-ERK and PI3K/Akt signaling pathways. a Heatmap showing the 72 downregulated genes in miR-199b-overexpressing H1299 cells. b Venn diagram showing candidate target genes of miR-199b in NSCLC. Candidate target genes of miR-199b were indicated using TargetScanHuman from the 72 genes that were downregulated by miR-199b overexpression in H1299 cells. c Sequence alignment of miR-199b with the 3`-UTR of the K-Ras, PIK3R1 (PI3K p85), Akt1, Rheb and KSR2 genes. d-e miR-199b negatively regulates the expression of the candidate target genes of miR-199b at both the mRNA and protein levels in NSCLC cells. The mRNA and protein expression levels of the indicated genes were measured using qRT-PCR and Western blot analysis in stably expressing miR-199b A549 cells and stably expressing miR-199b-antisense H522 cells. f The expression of candidate target genes of miR-199b was significantly decreased in lung tissues from miR-199b-delivered K-RasLA1 transgenic mice. The expression levels of the indicated proteins were detected in lung tissues of miR-199b-delivered K-RasLA1 transgenic mice and their control groups using Western blot analysis. g 3`-UTR luciferase reporter assay for candidate genes of miR-199b. HEK293 cells were cotransfected with miRNA luciferase reporter plasmid harboring indicated gene mutants or wild-type 3’UTR and miR-199b mimics or negative oligonucleotides control (NC). After 48 h of transfection, luciferase intensity was assessed

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