Bel 7404

Human HCC cell lines, SMMC-7721 and BEL-7404 (7721 and 7404), were obtained from the Cell
Bank of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, cultured
in RPMI1640 medium supplemented with 10% fetal bovine serum, 100 μg/ml penicillin, and 100
μg/ml streptomycin, and maintained in an incubator with a humidified atmosphere of 5%
CO₂ at 37°C. For B[a]P exposure, 7404 cells were co-cultured with 0.01, 1,
and 100 nM B[a]P or 0.1% DMSO for up to 4 weeks. After treatment, B[a]P was withdrawn and
the effects of B[a]P on HCC cells were determined. Cell morphology was observed using an
inverted microscope. CCK-8 (Dojindo) was used to measure cell growth. miRNA mimics and
inhibitors were purchased from Gene Operation. B[a]P, propidium iodide, crystal violet for
migration staining, and other chemicals used in this study were purchased from
Sigma-Aldrich.

The HCC cell lines, MHCC97H, MHCC97L, SMMC-7721, BEL-7404, and the normal hepatocyte lines, WRL-68 and L02, were purchased from the Shanghai Cell Bank (Shanghai Institute for Biological Science, Chinese Academy of Science, Shanghai, China). All cell lines were cultured in media according to the provider's instruction. The preconditioned cells used in the in vitro experiments were cultured in media containing 20 ng/ml bFGF (Sigma-Aldrich, Shanghai, China), or in media containing 0 to 1,000 nmol/L rapamycin (Sigma-Aldrich, Shanghai, China). Cells were also preconditioned by infection of adenovirus (Ad5-hSulf1, Ad5-EGFP) [13 (link)] at a multiplicity of infection (MOI) of 100 pfu/cell. All the parental and preconditioned cells were cultured for 48 h, and harvested.

The human tumor cell lines (BEL-7404, HepG2, MGC80–3, T-24, SK-OV-3/DDP and HeLa cells) and the human normal liver cell line HL-7702 were obtained from Shanghai Cell Bank in Chinese Academy of Sciences. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) or RPMI-1640 medium, which supplemented with 10% fetal bovine serum (FBS) and 100 U/mL penicillin and streptomycin. Cells were cultured at 37 °C in a humidified atmosphere with 5% CO₂/95% air. Tested compounds (2.0 mM) were prepared as DMSO stock solution before they were diluted to working concentrations by PBS buffer (containing 0.5% DMSO), while cisplatin dissolved in 0.9% sodium chloride solution was used as positive controls.

Human liver cancer cells (BEL-7404, Hep3B, Huh-7, and HepG2) and human liver cell L02 were obtained from Shanghai Cell Bank (Shanghai, China) or ATCC (the American Type Culture Collection). All the cell lines used in this study were approved by the Ethics Committee of the First Affiliated Hospital of Nanchang University. All cell lines were cultured in 90% DMEM and 10% FBS at 37°C in a 5% CO₂ incubator humidified atmosphere as recommended.

Hepatoma cell lines Hep3B, Huh-7, HepG2, SNU-449, SMCC-7721, MHCC-97h, BEL-7404, and liver normal epithelial cells L-02 were purchased from Shanghai Cell Bank, Chinese Academy of Sciences. Cells were maintained in culture medium with 10% fetal bovine serum (Clark) and placed in an incubator with 5% carbon dioxide at 37°C. Both DMEM and RPMI-1640 medium were purchased from Gibco Company. CCK-8 kits were purchased from Beyotime Company. Quercetin reagent was obtained from Sigma Company. The anti-Nosip antibody was obtained from Proteintech Company, while anti-β-actin antibody was purchased from Abclonal Company.

MHCC97L, MHCC97H, HCCLM3, HCCLM1-S3, HCCLM1-S4, HCCLM1-S5, HCCLM1-S11, HCCLnM1-S11, HCCLnM1-S13 were established in our lab [11 (link)–13 (link)], CSQT-1, CSQT-2, SMMC-7721 were gifted from Second Military Medical University, Shanghai, China, Bel-7402, Bel-7404 were obtained from the Shanghai Cell Bank, Chinese Academy of Sciences. Other cells were purchased from the American Type Culture Collection. The HCC cell line Hep3B was cultured in RPMI-1640 medium with 10% fetal bovine serum (Hyclone, USA) and 1% penicillin-streptomycin (Invitrogen, USA). Other HCC cells were grown in DMEM medium (Hyclone, USA) with 10% fetal bovine serum and 1% penicillin-streptomycin. All cell lines were maintained in the incubator at 37 °C at 5% CO₂. The metastatic status and evolutionary relationship of these cell lines were listed in Additional file 1: Table S1.