Mhcc97h

Hepatocellular carcinoma cell lines, including Bel-7402 (catalogue number TCHu 10), QGY-7703 (catalogue number TCHu 43), MHCC97L (catalogue number CC0109), MHCC97H (catalogue number SCSP 528) were cultured in RPMI 1640 (Gibco BRL, Rockville, MD) supplemented with 10% fetal bovine serum (Gibco BRL, Rockville, MD), penicillin (100 units/ml) and streptomycin (100 units/ml), and maintained in a 5% CO₂-humidified incubator at 37 °C. All cell lines were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China).

Human HCC cell lines Huh7, MHCC97H, MHCC97L, HCCLM3, HepG2, Hep3B, and human embryonic kidney cells (293 T) were purchased from the American Type Culture Collection (ATCC). MHCC97H, MHCC97L, HCCLM3, HepG2, Huh7, and 293 T cell lines were cultured in DMEM medium (Gibco, CP21110161839) containing 10% fetal bovine serum (FBS) (Gibco, 10099141C) and 1% penicillin–streptomycin solution (Gibco, 15140112). The MEM medium (Gibco, 8122107) containing 10% FBS and 1% penicillin–streptomycin solution was used to culture Hep3B cells. All cell lines were cultured in a humid environment (5% CO2, 37°C).

Human HCC cell lines HEP3B and SNU387 were purchased from ATCC. Human HCC cell lines Huh7 and MHCC-97H were purchased from the National Collection of Authenticated Cell Cultures (https://www.cellbank.org.cn/). HEP3B cells were cultured in Minimum Essential Media (MEM, Gibco, 11095080) Supplementaryemented with 1% Non-Essential Amino Acids (NEAA, Sigma Aldrich, M7145) and 10% fetal bovine serum (FBS, Gibco, 12662029). Huh7, MHCC-97H, and SNU387 cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, 11995065) with 10% fetal bovine serum (FBS, Gibco, 12662029). All cells were cultured in a humidified incubator containing 5% CO₂ at 37 °C.

The human intrahepatic bile duct epithelial cells (HIBEpiCs) were purchased from ICell Bioscience Inc. (Shanghai, China). The human hepatocellular carcinoma cell line MHCC-97h and human mononuclear cell line THP-1 were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). The HIBEpiC and THP-1 were cultured in RPMI-1640 medium (Gibco, Grand Island, USA), and MHCC-97h was cultured in DMEM medium (Gibco, Grand Island, USA), all media containing 10% fetal bovine serum (Gibco, Grand Island, USA) and 1% penicillin/streptomycin (Beyotime, Shanghai, China). All three cell lines were grown in an incubator (37°C, 5% CO₂).
Cobalt chloride (CoCl₂, Sigma-Aldrich, USA) was used to simulate the hypoxia process of HIBEpiCs. The HIBEpiCs were divided into four groups: the control group (CON), the IL-22 (10 ng/ml) (Recombinant human IL-22, Absin, Shanghai, China) treatment group, the CoCl₂ (150 μM) treatment group, and the CoCl₂ (150 μM)+IL-22 (10 ng/ml) treatment group.

THLE-2, MIHA, Hep3B and Huh7 cells were purchased from the National Collection of Authenticated Cell Cultures (https://www.cellbank.org.cn/). SNU-449, SNU-182, and SNU-387 were purchased from ATCC: The Global Bioresource Center (ATCC, https://www.atcc.org/). PLC/PRF/5 and MHCC-97H were provided by Prof. Lu Guo-dong of the School of Public Health, Guangxi Medical University. Hep3B was cultured in MEM medium (Gibco, Shanghai, China) supplemented with 10% FBS (Gibco, Shanghai, China). THLE-2 was cultured in BEGM medium (Lonza, CC3170, Hong Kong). SNU-449, SNU-182, and SNU-387 were cultured in RMPI-1640 medium (Gibco, Shanghai, China) supplemented with 10% FBS (Gibco, Shanghai, China). MIHA, LM3, Huh7, PLC5, and MHCC97-H were cultured in DMEM medium (Gibco, Shanghai, China) supplemented with 10% FBS (Gibco, Shanghai, China).

The human liver cell line LO2 and hepatocellular cell lines SNU398, SNU449, MHCC97H, Hep3B, PLC/PRF/5, and HepG2 were purchased from the ATCC (Manassas, VA, United States). PLC/PRF/5, Hep3B, MHCC97H, and HepG2 cell lines were cultured in complete Dulbecco’s modified Eagle’s medium (Gibco, United States), and SNU398 and SNU449 cells were cultured in RPMI1640 medium (Gibco, United States); all cells were supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified incubator with 5% CO₂ in the air.
HCC cell lines (Hep3B, HepG2, and SNU398) were transfected with siRNAs targeting ALG3 or control siRNA (HIPPOBIO, China) using Lipofectamine 2000 Reagent (Invitrogen, United States). The efficiency of the knockdown was confirmed using qRT-PCR analysis. The siRNA used were as follows: si-NC: 5′-UUC​UUC​GAA​GGU​GUC​ACG​UTT-3′, ALG3 siRNA#1: 5′-GGU​UUC​GUG​UAC​AUC​UUU​AUG-3′, and ALG3 siRNA#2: 5′-GGA​CCU​GAG​UCU​ACC​CUC​AGG-3’.

Human HCC cell lines MHCC-97H, HUH7, Bel-7402, Hep3B, HepG2 and SMMC-7721 were purchased from the cell bank of the Committee on Type Culture Collection of the Chinese Academy of Sciences (CTCC, Shanghai, China). THLE-2 and SNU449 were obtained from ATCC. MHCC-97H, SNU449, and MHCC-97L cell lines were maintained in RPMI 1640 medium (GIBCO, USA). HEK293T, HUH7, Bel-7402, Hep3B, HepG2 and SMMC-7721cells were cultured in DMEM (GIBCO, USA). Cell lines were maintained in culture supplemented with 10% FBS (GIBCO, USA) and 1% penicillin/streptomycin (Thermo) at 37°C with 5% CO₂ in a humidified incubator (Thermo).