Cell proliferation assay

TE-1 and KYSE150 were cultured in 96-well plates at 5 × 10³ in serum starvation overnight. Each well was handled with 10 μL reagents from Cell Counting Kit-8 (Beyotime Biotechnology, Shanghai, China) and then each well was supplemented with 10 μL CCK-8 and incubated at 37°C for 2 h. Three auxiliary holes in each group. Optical density (OD) was measured at 450 nm using a microplate reader (BioTek, USA) [43]. The cells were transfected by shRNA for 48 hours and then counted.