Hepg2

The human hepatoma cell line, HepG2, was obtained from Cell Resource Center, IBMS, CAMS/PUMC (Beijing, China) and cultured in Dulbecco’s Modified Eagle’s medium DMEM (DMEM, Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA), 1% non-essential amino acids (NEAA) (Life technologies, Carlsbad, CA, USA) and 1% penicillin–streptomycin at 37 °C, 5% (v/v) CO2. HepG2 cells were serum-starved for 18 h and then treated with liraglutide (Novo Nordisk, Bagsværd, Denmark) at various concentrations (0, 10, 50, 100, 500 and 1000 nM) for 24 h or with 500 nM liraglutide for different times (0, 3, 6, 12, 24 h).

The human carcinoma cell (HCC) lines HepG2, SMMC7721 and BEL7402 were maintained in DMEM medium (Gibco, Paisley, UK) supplemented with 10% fetal calf serum (FCS, Gibco, Paisley, UK) in humidified 5% CO2/95% atmosphere at 37°C. HepG2 (human hepatic carcinoma), SMMC 7721 (human hepatic carcinoma) and BEL7402 (human hepatic carcinoma) were purchased from Cell Resource Center, IBMS, CAMS/PUMC).

Cytotoxicity was tested by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method according to our previous report^{20 (link)}. Human cervical carcinoma cells (HeLa), human lung cancer cells (A549), and human hepatoma cells (HepG2) were purchased from Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and the cell lines were grown in RPMI-1640 culture medium with 200 μL/mL foetal bovine serum (FBS) under a humidified atmosphere of 5% CO₂ and 95% air at 37 °C. A 100 μL cell suspension at a density of 1.5 × 10⁵ cell mL⁻¹ was pipetted into 96-well microtiter plates. Fungal extracts with different concentrations from 100 to 600 μg/mL were added to each well and incubated for 24 h under the above conditions in a CO₂ incubator. Then, 20 μL of MTT (5 mg/mL) was added to each well, and the plates were further incubated for 3 h. DMSO (200 μL) was added to dissolve the formazan crystals. The absorbance was then measured at 570 nm by a microplate reader. The cell inhibition rate (%IR) was calculated by the following equation: % IR = [(A_bla − A_sam)/A_bla] × 100, where A_bla is the absorbance of the blank and A_sam is the absorbance of the test compounds. The IC₅₀ value was calculated from the dose–response relationship. Doxorubicin was used as the positive control.