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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: KLF5-induced BBOX1-AS1 contributes to cell malignant phenotypes in non-small cell lung cancer via sponging miR-27a-5p to up-regulate MELK and activate FAK signaling pathway

Fig. 4

BBOX1-AS1 interacts with miR-27a-5p to suppress its expression. a lncLocator was used to predict the subcellular localization of BBOX1-AS1. b Subcellular fractionation assays were used to confirm the cytoplasm and nucleus distribution of BBOX1-AS1 in NSCLC cells. c Among the down-regulated miRNAs in NSCLC from GSE135918, three are found to be able to potentially interact with BBOX1-AS1 according to DIANA-LncBase and LncBook. d GSE135918 showed the expression of miR-27a-5p, miR-3664-3p and miR-4778-3p in 5 tumor tissues and matched normal tissues from NSCLC patients. e Dual-luciferase reporter assays were performed to detemine the influence of miR-27a-5p, miR-3664-3p and miR-4778-3p on the luciferase activity of BBOX1-AS1-wt reporter in HEK293T cells. f Schematic representation of wt- and mut-BBOX1-AS1 binding sequences on miR-27a-5p. Red fonts represented the mutant bases. g Dual-luciferase reporter assays were conducted in A549 and SK-MES-1 cells after transfection with BBOX1-AS1-wt or BBOX1-AS1-mut reporter and miR-NC or miR-27a-5p. h RNA pull-down assays were applied to examine the binding between miR-27a-5p and BBOX1-AS1 in NSCLC cells by using Biotin-NC or Biotin-miR-27a-5p. i The expression of miR-27a-5p was detected by qRT-PCR assays in NSCLC cells with BBOX1-AS1 overexpression or depletion. j qRT-PCR analysis of miR-27a-5p expression in NSCLC tumor tissues and corresponding non-cancerous. k Correlation analysis between miR-27a-5p and BBOX1-AS1 in NSCLC tumor specimens. ***P < 0.001

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