Evaluating Cell Viability and Proliferation

The cell viability of HCC cells was evaluated by the MTT assay. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT, Roche, USA) was used for the MTT assay as described previously^{16 (link)}. EdU and colony formation assays were performed for the evaluation of cell proliferation. For the EdU assay, a cell proliferation ELISA, the Cell-Light™ EdU Apollo®488 In Vitro Imaging Kit (#C10310-3, RiboBio Co., LTD, Guangzhou, China), was used as previously described^{20 (link)}. For the colony formation assay, HCC cells (1 × 10³) were seeded in 6-well plates and cultured for 2 weeks in DMEM medium with 10% FBS. The cell colonies containing more than 30 cells were counted as single colonies^{20 (link)}.