Human LUSC NCI-H226/SK-MES-1 cells were planted in 96-well plates at a density of 3000 cells/well and cultured for 24 h. Thereafter, the cells were transfected with miR-665 mimics, control, miR-665 inhibitor, negative control (NC), TRIM8 siRNA (60 nM), negative control siRNA (NC-siRNA, 60 nM), TRIM8 expression plasmid or control vector for 24, 48, 72 or 96 h, respectively. Each well was incubated with 10 μg Cell Counting Kit-8 solution (Dojindo, Shanghai, China) for 4 h. Cell proliferation was estimated by examining the absorbance at 450 nm with Varioskan Flash Spectral Scanning Multimode Reader (Waltham, MA, USA).

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