Mechanism of miR-548b-5p down-regulating FZD7 to impede the migratory and invasive behavior of gastric carcinoma cells.

Abstract

This study aimed to investigate the possible mechanism of Micro RNA-548b-5p (miR-548b-5p) down-regulating frizzled (FZD) 7 to suppress the migration and invasion of gastric carcinoma cells. For this purpose, HGCC (Human gastric carcinoma cell) lines were selected (Hs-746T, NCI-N87, SGC-7901, MKN-45, SNU-1), and human normal gastric mucosa cells GES-1. QRT PCR was adopted to reveal and screen the cell line with low expression of mir-548b-5p (hs-746t) for research; the Hs-746T cells were randomly assigned into control group, miR-548b-5p NC group, miR-548b-5p mimic group, miR-548b-5p mimic+pc-FZD7 group. The CCK-8 assay was utilized to measure Hs-746T cell viability, while flow cytometry, Trans well chamber, and scratch test were utilized to examine the apoptotic, invasive, and migratory properties of the cells, respectively. WB was used to detect the SATB1, as well as the expression levels of proteins involved in apoptosis, including Caspase-3, Bax, and Bcl-2, as well as Matrix metalloproteinase 2 and 9 (MMP-2 and MMP-9) in SW620 cells. The binding of miR-548b-5p to FZD7 was evaluated through the dual-luciferase reporter assay. The results indicate that MiR-548b-5p showed low expression in HGCCs; in contrast to the control group (P>0.05), the Hs-746T cell viability, invasion, migration ability, MMP-2, MMP-9 protein significantly downregulated in miR-548b-5p mimic group (P<0.05), the apoptosis rate, Caspase-3, Bax protein expression were upregulated markedly, and Bcl-2 protein expression was downregulated significantly (P<0.05); in contrast to miR-548b-5p mimic group, the Hs-746T cell viability, invasion, migration ability, MMP-2, MMP-9 protein significantly were upregulated in miR-548b-5p mimic+pc-FZD7 group (P<0.05), the apoptosis rate, Caspase-3, Bax protein expression were significantly, and the level of Bcl-2 was down-regulated significantly (P<0.05); Double Luciferase Report shows that mir-548b-5p can target and regulate fzd7. It was concluded that MiR-548b-5p can suppress cell growth and migration of HGCC Hs-746T, which may be achieved by targeted down-regulation of FZD7.