Hcclm3 cells

The HCCLM3 cells (human hepatocarcinoma cell line) were received from the China Center for Type Culture Collection (CCTCC, Wuhan, China). The HCCLM3 cells were cultured in RPMI-1640 medium (Invitrogen) with 10% fetal bovine serum (FBS). Cells were incubated at 37°C in an incubator with 5% CO₂. Cells were passaged when they reached 70–80% confluence.

The HCCLM3 cells were purchased from the China Center for Type Culture Collection (CCTCC), and the Huh7 cells were purchased from Suyan Biotechnology Co., Ltd. The THLE-2 cells were purchased from Meisen Cell Technology Co., Ltd. The HCCLM3 and Huh7 Cells were cultured in DMEM/high-glucose medium (Gibco; Thermo Fisher Scientific, Inc.) with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin (Gibco; Thermo Fisher Scientific, Inc.) at 37°C and 5% CO₂. The THLE-2 cells require a special coating medium (CTCC-002-30, Meisen Cell Technology Co., Ltd.) and were incubated overnight in an incubator at 37°C and 5% CO₂ and cultured in BEGM™ BulletKit™ medium (cat. no. CC-3170, Lonza Group, Ltd.) with 10% FBS. The HCC cells were observed and representative images were captured under an inverted microscope (Ts2R-FL, Nikon Corporation; ×100 magnification) at 0 and 24 h following drug intervention. The HCC cells were treated with PPVI (4 µM) at 37°C for 24 h, following which different assays were performed. Inhibitors, such as 3-MA (1 mM), Ac-DEVD-CHO (50 µM) and Fer-1 (1 µM) for 2 h prior to treatment with PPVI (4 µM) for 24 h.