Cell migration and invasion capabilities were detected using Transwell chambers (Corning, NY, USA) either uncoated or coated with Matrigel matrix (BD, Science, Sparks, MD, USA) as previously described [10 ].
Cell Titer 96 nonradioactive cell proliferation (MTS) (Promega BioSciences, Madison, WI, USA), Cell‐Light™ EdU cell proliferation detection (EdU) (RiboBio), and colony formation assays were performed to test the proliferation ability of the GC cells following the manufacturer's protocols. For the MTS assay, cell proliferation was measured at 24, 48, and 72 h after transfection. MTS (5 mg·mL−1) was added to the culture medium, followed by an incubation of 2 h, and absorbances were read at a wavelength of 490 nm. For the EdU assay, after EdU incubation, the cells were treated with an Apollo reaction cocktail, stained with DAPI, and visualized under a fluorescent microscope (Olympus, Tokyo, Japan). For the colony formation assay, 500 cells were seeded into six‐well plates. After incubation for 2 weeks, the colonies were fixed and stained, and then, formative clones were counted.
Cell Titer 96 nonradioactive cell proliferation (MTS) (Promega BioSciences, Madison, WI, USA), Cell‐Light™ EdU cell proliferation detection (EdU) (RiboBio), and colony formation assays were performed to test the proliferation ability of the GC cells following the manufacturer's protocols. For the MTS assay, cell proliferation was measured at 24, 48, and 72 h after transfection. MTS (5 mg·mL−1) was added to the culture medium, followed by an incubation of 2 h, and absorbances were read at a wavelength of 490 nm. For the EdU assay, after EdU incubation, the cells were treated with an Apollo reaction cocktail, stained with DAPI, and visualized under a fluorescent microscope (Olympus, Tokyo, Japan). For the colony formation assay, 500 cells were seeded into six‐well plates. After incubation for 2 weeks, the colonies were fixed and stained, and then, formative clones were counted.