Journal: Translational Oncology

Article Title: MiR-196b-5p activates NF-κB signaling in non-small cell lung cancer by directly targeting NFKBIA

doi: 10.1016/j.tranon.2023.101755

Figure Lengend Snippet: NFKBIA is a direct target of miR-196b-5p. (A) Western blot analysis of QKI-5 in QKI-5 knockdown H1299 cells and control cells. GAPDH was used as an internal control. (B) Lysates from QKI-5 knockdown H1299 cells or control cells were subjected to RIP analysis. The cell extracts were subjected to immunoprecipitation with IgG or anti-QKI5 antibody. Pull-down RNA was analyzed by qRT-PCR using specific probe for miR-196b. Data are presented as mean ± SD as determined by triple assays. (C) Schematic diagram showing matched sequences between the seed sequence of miR-196b-5p and 3′UTRs of NFKBIA. The arrows indicate the mutagenesis nucleotides. (D) Luciferase reporter constructs containing wild-type, mutated form of NFKBIA 3′UTR, or empty vector were co-transfected with pre-miR-196b precursor into the 293T cells. Data are presented as mean ± SD as determined by triple assays. The p -values were calculated by one-way ANOVA with Tukey's multiple comparisons test. WT, wild-type; Mut, mutated form.

Article Snippet: Antibodies against Ki67 (ab16667), CD31 (ab28364), and QKI-5 (ab232502) were purchased from Abcam (Cambridge, USA).

Techniques: Western Blot, Immunoprecipitation, Quantitative RT-PCR, Sequencing, Mutagenesis, Luciferase, Construct, Plasmid Preparation, Transfection

Journal: Translational Oncology

Article Title: MiR-196b-5p activates NF-κB signaling in non-small cell lung cancer by directly targeting NFKBIA

doi: 10.1016/j.tranon.2023.101755

Figure Lengend Snippet: Enhanced miR-224 is responsible for downregulated QKI-5 expression in NSCLC. (A) qRT-PCR measure miR-224 expression after overexpressing premiR-224 in H1299 and A549 cells. (B, C) Western blot analyses measure QKI-5 (B) and NFKBIA (C) protein expressions in H1299 and A549 cells after transfection with premiR-224 or control. (D) QKI expression from TCGA RNA-seq data and miR-224 expression from TCGA miR-seq data were used to examine the correlation between QKI and miR-224 expressions in LUSC dataset ( n = 235). (E) Schematic Figure of proposed miR-196b-5p mediated down-regulation of NFKBIA increases pSTAT3 expression by activating NF-κB signaling. The p -values in (A) were calculated by unpaired t -test.

Article Snippet: Antibodies against Ki67 (ab16667), CD31 (ab28364), and QKI-5 (ab232502) were purchased from Abcam (Cambridge, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, RNA Sequencing Assay