miR-374b-5p suppresses RECK expression and promotes gastric cancer cell invasion and metastasis

Figure 1 Differential expression of microRNAs in gastric cancer cells. A: microRNA (miRNA) expression in gastric cancer cell lines SGC-7901 and MGC-803 was compared with the normal GES-1 cell line using cluster analysis. All cell samples were detected in triplicate, such that the nine lanes include GES-1 (G1-G3), SGC-7901 (S1-S3) and MGC-803 (M1-M3). The color scale denotes the extent of relative upregulation (red) and downregulation (green); B: Histogram of miR-374b-5p expression detected by cluster analysis; C: Confirmation of miR-374b-5p expression via quantitative real-time reverse transcription-polymerase chain reaction (n = 3); ^aP < 0.05 vs GSE-1.

Figure 2 Effects of miR-374b-5p on MGC-803 cell metastasis and invasion. MGC-803 cells were transfected with a miR-374b-5p antisense oligonucleotide or negative control-oligo vector, or treated with phosphate-buffered saline (untreated), or transfection reagent only. A: Cells were subjected to a transwell invasion assay and detected at 48 h after seeding (left: Coomassie Brilliant Blue staining, × 200; right, quantification); B: Wound margin distance was measured after 48 h in a scratch assay (Coomassie Brilliant Blue staining, × 40); C: Absorbances in a cell counting kit-8 assay 96 h after transfection; (n = 3), ^aP < 0.05 vs Anti-miR-374b-5p.

Figure 3 Regulation of RECK expression in MGC-803 cells by miR-374b-5p. A: The miR-374b-5p binding site within the 3′ UTR of human (hsa) RECK cDNA was predicted with PicTar, TargetScan and miRanda software; B: Cells were co-transfected with RECK 3’UTR-pmirGLO and a plasmid containing either miR-374b-5p or miR-NC plasmid, ^aP < 0.05 vs miR-NC (n = 3); C: Western blot of proteins; D: qRT-PCR from cells transfected for 48 h with anti-miR-374b-5p or a negative control vector, or treated with phosphate-buffered saline (untreated), or transfection reagent only; (n = 3), ^cP < 0.05 vs Anti-miR-374b-5p.