11. MicroRNA-129-2-3p regulates cell proliferation by targeting Gli1 in gastric cancer

Objective: The purpose of this study was to investigate the regulation of expression of Gli1 in microRNA level in gastric cancer.
Methods: Target miRNA were predicted based on the intersection of three public databases and our previous work. Gli1 regulated by miR-129-2-3p was detected using Dual Luciferase Reporter system. The protein expression of Gli1 regulated by miR-129-2-3p was detected by western blotting. We screened the expressing levels of miR-129-2-3p and Gli1 in gastric cancer cell lines. We transfected miR-129-2-3p mimics into MNK-28 cell to observe the change of ability of colony formation and cell growth in vitro. The effect of miR-129-2-3p to tumorigenicity in vivo was studied by an experiment in nude mice.
Results: Using a luciferase activity assay and western blot, Gli1 was targeted by miR-129-2-3p in cells. Western blotting also showed miR-129-2-3p can downregulate the protein expression of Gli1 in gastric cancer cells. We showed significant down-regulation of miR-129-2-3p in 80% gastric cancer tissues detected, compared with their nontumor counterparts by realtime RT-PCR. We also found that miR-129-2-3p could inhibit cell proliferation in vitro by MTT assay, growth curves and colony formation in MNK-28 cells. An experiment in nude mice revealed that.