Mhcc 97l

Immortalized human hepatocyte L02, and the HCC cell lines SMMC7721, HepG2, MHCC97L, HCCLM3, Huh7 and Hep3B were purchased from the China Center for Type Culture Collection (Wuhan, China). YY8103 cells were obtained from the Key Laboratory on Living Donor Liver Transplantation. All the cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen, Thermo Fisher Scientific, Inc., Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, Life Technologies, Carlsbad, CA, USA), 50 U/ml penicillin (Invitrogen) and 50 U/ml streptomycin (Invitrogen) in a humidified 5% CO2 incubator at 37°C.

The indicated hepatocellular carcinoma cell lines (SMMC-7721, HCC-LM3, Huh7, HepG2, MHCC-97 L, MHCC-97H) and normal liver cell line (THLE-3) were obtained from the China Center for Type Culture Collection (Wuhan, China) and were authenticated by the provider using DNA-fingerprinting or isoenzyme analysis. All cell lines were maintained in DMEM (HyClone, UT, USA) containing 10% fetal bovine serum and 1% penicillin/streptomycin (GibcoBRL, MD, USA) at 37°C in a humidified atmosphere of 5% CO2.

Human low metastatic liver cell MHCC-97L was a gift from Prof. Fubing Wang from the Hubei Key Laboratory of Tumor Biological Behaviors of the Zhongnan Hospital of Wuhan University School of Medicine, and high metastatic liver cell HCC-LM3 was purchased from the China Center for Type Culture Collection (CCTCC) of Wuhan University, China. Human hepatocellular carcinoma cells Huh7.5.1, normal liver cells L02, MHCC-97L, and HCC-LM3 were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS, Gibco) (Invitrogen, USA) at 37°C in a 5% CO₂ atmosphere as previously described (20 (link), 21 (link)).

A total of 20 HCC tissues and the corresponding adjacent noncancerous tissues were obtained from Department of Hepatobiliary Surgery, Xinqiao Hospital, Third Military Medical University (Chongqing, China). Fresh tissue samples were collected and snap frozen in liquid nitrogen. The study was approved by the ethics committee of Third Military Medical University (Chongqing, China).
Human HCC cell lines HepG2 and Hep3B were from American Type Culture Collection (Manassas, VA). The other HCC cell lines including Huh7, PLC, SMMC-7721, MHCC97L and MHCC97H, and hepatic cell line L02 were purchased from China Center for Type Culture Collection (Wuhan, China). All cells were cultured in Dulbecco’s Modified Eagle’s medium supplemented with 10 % fetal bovine serum, streptomycin (100 mg/mL) and penicillin (100 U/mL) at 37 °C in a 5 % CO₂ humid incubator.

One human normal hepatic epithelial cell line THLE-21 and six HCC cell lines (Hep3B, Huh7, MHCC97L, SK-hep1, SNU-387, SMMC7721) were purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS) (Gibco, USA) in a humidified 37 °C incubator with 5% CO₂. All cell lines used in our study have been authenticated using STR profiling. For circTP63 or ZBTB18 overexpression, the full-length sequence of circTP63 or ZBTB18 was cloned into pcDNA3.1 (Invitrogen, CA, USA) plasmid to generate pcDNA3.1-circTP63 or pcDNA3.1-ZBTB18. All small interfering RNAs (siRNAs), miR-155-5p mimics, inhibitor, and their NC negative controls were obtained from GeneCopoeia (Guangdong, China). Cell transfection was conducted with Lipofectamine 2000 Reagent (Invitrogen, CA, USA) as the manufacturer’s protocol. The siRNA sequences for transfection as follows:
si-circTP63#1, 5′-GCCAACAGUGAGGGGCCGU-3′;
si-circTP63#2, 5′-CAACAGUGAGGGGCCGUGAGA-3′;
siRNA control (si-NC) siRNAs 5′-UUCUCCGAACGUGUCACGU-3′.