Matrigel transwell

CCK-8 assay was used to detect the cell proliferation, the cells (2000) were seeded into 96-wells plates. The next day, the cells were transfected with PSMA1 siRNA or PSMA1 overexpression plasmid. Then, cell viability was detected by a CCK-8 (Solarbio Life Science, Beijing, China) assay at 450 nm. The colony formation assay was performed to detect the cell proliferation, and the cells (800) were plated in 6-wells in each group and cultured. After 2 weeks, cell colonies in the plates were fixed with 4% paraformaldehyde and stained with 1% crystal violet. The numbers of cell colonies were counted using Image J. The transwell assay was used to detect the migration and invasion abilities of gastric cancer cells and evaluated by coating with or without Matrigel Transwell (Corning, USA). 1.5 × 10⁴ AGS cells and 3 × 10⁴ BGC-823 cells in 200 μl serum-free medium were seed into the upper chamber with or without pre-coated Matrigel, and containing 20% FBS medium was moved into the lower chamber. After 48 h, the migratory or invasive cells were fixed with 4% paraformaldehyde for 20 min and stained with 1% crystal violet for 30 min. The number of invading cells was counted in ten random visual fields per chamber under a microscope (Olympus, Tokyo, Japan).

Twenty-four-well Matrigel Transwell (Corning, NY, United States) was used to investigate cell invasion; 2 × 10⁵ TCCSUP or SW780 was seeded on precoated Matrigel (1 μg/μL, BD Biosciences, CA, United States). Medium including fetal bovine serum was used to stimulate invasion in the bottom of wells. After 48 h, the invasion cells were stained by 0.1% crystal violet solution.