Mhcc 97h

HEK293T cells were obtained from the American Type Culture Collection (ATCC). Human hepatocellular carcinoma (HCC) cell lines, including Huh7, LM3, Hep3B, MHCC97H and HepG2,and the mouse HCC cell line Hep1-6 were purchased from National Collection of Authenticated Cell Cultures. All these cell lines were cultured in high-sugar Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), streptomycin (100 μg/mL), and penicillin (100 U/mL). Working cultures were maintained at 37°C in a humidified incubator with 5% CO2.

Matched pairs of cancerous and normal tissue from 50 patients with HCC were obtained from the Second Affiliated Hospital of Nantong University. The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). This study was approved by Ethics Committee of the Second Affiliated Hospital of Nantong University (No. 2020KT014). The study mainly uses biological specimens obtained in previous clinical diagnosis and treatment, and is a secondary use of biological specimens, which is approved by the ethics committee without informed consent. The clinical characteristics of the HCC patients are summarized in Table 1.
The human HCC cell lines (Bel-7404, HCCLM3, MHCC-97H, Hep G2, and Huh-7) and normal human hepatocytes LO2 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All cell lines were cultured in Dulbecco’s Modified Eagle Medium (DMEM) medium (product code: 11960044; Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS) (product code: 10099141C; Corning, Corning, NY, USA) and 1% penicillin and streptomycin (product code: 15140122; Gibco, Thermo Fisher Scientific, Grand Island, NY, USA) in a humidified incubator at 37 ℃ with a 5% CO₂ atmosphere.

Hepatocellular carcinoma cell lines MHCC‐97H, HCC‐LM3, MHCC‐97 L, HepG2, Huh‐7, PLC, SMMC‐7721, Bel‐7402, and Hep3B were obtained from the cell bank of the Chinese Academy of Sciences (Shanghai, China). Short tandem repeat (STR) profiling of HCC cells was verified at GenePharma (Shanghai, China). All cell lines were maintained under standard conditions in high glucose DMEM (Thermo Fisher) with 10% FBS. MG132 (SML1135), CoCl₂ (232696), DM‐2OG (D3695), D‐2‐HG (H8378), and L‐2‐HG (90790) were obtained from Sigma.

The HCC cell lines HepG2, MHCC-97L, MHCC-97H, Huh7, and LM3 and the normal liver cell line L02 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Hep3B cells were obtained from the American Type Culture Collection (Manassas, VA, USA). All cell lines were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, USA) supplemented with 10% fetal bovine serum (FBS, Gibco USA), 100 U/mL penicillin, and 100 U/mL streptomycin and incubated at 37 °C under an atmosphere containing 5% CO₂.