Evaluating PVT1 Knockdown Effects

Cells were grown in 96-well plates and transfected with siRNA (siPVT1-1/2). Subsequently, a Cell Counting Kit-8 (CCK-8, Dojindo, Shanghai, China) was used to detect cell proliferation at 0, 24, 48, 72, 96 and 120 h following the manufacturer’s protocol. For Transwell invasion assays, cells were inoculated into Transwell plates containing Matrigel. DMEM with 20% FBS was placed in the lower chamber. Serum-free DMEM was used to resuspend the cells, which were added to the upper chamber. After 24 h, 70% methanol was used to fix the membranes, which were then stained using 0.1% crystal violet for 10 min. After thorough washing with PBS, the membranes were imaged. The average number of invasive cells was determined by counting in three randomly chosen fields under a microscope.