Mhcc97l

Nineteen paired HCC and adjacent non-tumor liver tissues were collected from patients who underwent surgical resection at the Third Affiliated Hospital of Sun Yat-sen University in Guangzhou, China. The paired non-tumor tissues were collected at least 2 cm away from the primary HCC. Both tumor and non-tumor tissues were histologically confirmed. All patients were unrelated ethnic Han Chinese who lived in Southeast China. None of the patients had received local or systemic anticancer treatments prior to the operation. This study was approved by the Institute Research Ethics Committee at the Third Affiliated Hospital of Sun Yat-sen University. Informed consent was obtained from each patient.
Cell lines derived from human hepatocellular carcinoma (MHCC97L, SMMC-7721, QGY-7703, BEL-7402, and BEL-7404) were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China), and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM, Life Technologies, Grand Island, NY, United States) supplemented with 10% fetal bovine serum (FBS, Hyclone, Thermo Fisher Scientific, Victoria, Australia).

HL7702, Hep3B, MHCC97L and MHCC97H cells were obtained from the Cobioer Bioscience Company (Nanjing, China), and HCCLM3 cells were obtained from China Center for Type Culture Collection (Wuhan, China). HL7702 cells were cultured in 1640 medium (Life Technology, Carlsbad, CA, USA), and Hep3B, MHCC97L, MHCC97H and HCCLM3 cells were cultured in high‐glucose DMEM (Life Technologies Corporation, Carlsbad, CA, USA) and supplemented with 10% FBS (Life Technologies Corporation) in a humidified atmosphere of 5% CO₂. For GSK3β inhibition, MHCC97L and HCCLM3 cells were firstly incubated with 1 or 4 μg/ml BS‐I (Sigma‐Aldrich, St Louis, MO, USA) for 6 hrs, and then, 0.2 μM CHIR99021 (Selleckchem, Houston, TX, USA) or 4 mM LiCl (Sigma‐Aldrich) was added to the medium.

Human liver cancer cell lines (HepG2, Hep3B and Huh7) and human immortal liver cell lines (MIHA and LO₂) were purchased from the American Type Culture Collection (Manassas, VA, USA). The metastatic human liver cancer cell line MHCC97L was obtained from the Liver Cancer institute and Zhongshan Hospital of Fudan University, Shanghai, the People's Republic of China [39 (link)]. All the cell lines were cultured as previously described [40 (link)]. The pcDNA 3.1 (+) vector was purchased from Invitrogen (Carlsbad, CA). MHCC97L and Huh7 cells were transfected with recombinant plasmid using lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA) and cultured for two weeks under G418 selection. Stable clones were maintained in 0.05 mg/mL G418 for Huh7 cells and 0.5 mg/mL for MHCC97L cells, respectively.