Totally, 109 paired fresh HCC tissues (tumorous and adjacent normal samples) and relevant clinical information were obtained from patients undergoing hepatectomy at the First Affiliated Hospital of Nanjing Medical University (Nanjing, China) from June 2013 to July 2014. The study protocol was approved by the Institutional Ethics Committee of the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. All patients had given written informed consent to participate in our study before surgery and that was conducted in accordance with the Declaration of Helsinki. All fresh tissues were collected and immediately frozen in liquid nitrogen. The diagnosis of all patients was confirmed by pathology. HepG2, Hep3B, MHCC97H, Huh7, SNU423, and SMMC-7721 human hepatoma and normal L02 cell lines were obtained from Nanjing KeyGen Biotech Co. Ltd (Nanjing, China). All cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA) and 80 U/mL of penicillin sodium at 37°C in humidified air containing 5% carbon dioxide.
Hep3b
Manufactured by Keygen Biotech
Sourced in China
The Hep3B is a cell line derived from a human hepatocellular carcinoma. It is commonly used in research applications to study liver-related diseases and the biology of hepatocellular carcinoma.
Automatically generated - may contain errors
Lab products found in correlation
Hepg2, by Keygen Biotech (8 mentions)
Mhcc 97h, by Keygen Biotech (8 mentions)
Smmc 7721, by Keygen Biotech (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Snu423, by Keygen Biotech (4 mentions)
Mhcc 97l, by Keygen Biotech (4 mentions)
Sk hep 1, by Keygen Biotech (4 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Dmem medium, by Thermo Fisher Scientific (2 mentions)
Human normal l02 cell line, by Keygen Biotech (2 mentions)
Bel 7402, by Keygen Biotech (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Plc prf 5, by Keygen Biotech (1 mentions)
Thle 2, by Keygen Biotech (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Bel 7404, by Keygen Biotech (1 mentions)
Hcclm3, by Keygen Biotech (1 mentions)
12 protocols using hep3b
1
Comprehensive HCC Tissue Collection and Cell Lines
2
Hepatocellular Carcinoma Tissue and Cell Collection
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110 paired HCC fresh tissues consist of tumors and adjacent normal samples were obtained from patients who underwent liver resection at the Liver Transplantation Center in The First Affiliate Hospital of Nanjing Medical University between October 2014 and November 2015. All patients provided their written informed consent to participate in this study. The fresh tissue samples which were confirmed by the histopathological examination were collected in the operating room and processed immediately. Each sample was frozen and stored at liquid nitrogen. The HepG2, SNU423, SMMC-7721, Hep3B, 97H, 97L and Huh7 HCC cell lines and the human normal L02 cell line used in this study were obtained from KeyGen (Nanjing KeyGen Biotech Co.Ltd, China). All of the cells were cultured in DMEM medium(GIBCO, Carlsbad, USA) pre-treated with 10% fetal bovine serum, 80 U/ml of penicillin sodium at 37°C in humidified air containing 5% carbon dioxide.
3
Cell Culture Maintenance Protocol
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Hela, PLC-PRF-5, HepG2, MHCC-97L, MHCC-97H, LO2, THLE2, Hep3B, and SK-Hep-1 cells were purchased from KeyGEN BioTECH and OBIO, Shanghai. Cells are maintained in RPMI 1640 or DMEM medium (Neuronbc) with 10%FBS and 1%PS. All the cell lines were kept at 37 °C under a humidified atmosphere of 5% CO2 in an incubator, trypsinized, and passaged every 2 days. Plasmid DNA was transfected using Lipofectamine 2000 transfection reagent (Invitrogen).
4
Culturing Liver Cell Lines for Research
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The human normal liver cell line L-O2 and the HCC cell lines Huh7, Bel-7402, Bel7402/5FU, Hep3B, SK-Hep-1, and 97-H were obtained from KeyGen Biotech Co., Ltd. The cell lines were cultured in RPMI 1640 (Gibco, USA) or DMEM (Gibco, USA) supplemented with antibiotics (1× penicillin/streptomycin 100 U/ml, Gibco, USA) and 10% fetal bovine serum (Gibco, USA). Cells were incubated at 37°C in a humidified atmosphere containing 5% CO2.
5
Comprehensive Characterization of Liver Cancer Cell Lines
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The following human cell lines were used: SMMC-7721 (GCC-LI0007RT), Bel-7402 (GCC-LI0008RT), Bel-7404 (GCC-LI0005RT), Hep3B (GCC-LI0002RT), and Huh7 (GCC-LI0006RT) HCC cell lines, the hepatoblastoma cell line HepG2 (GCC-LI0003RT), the liver cancer cell line SK-Hep-1 (GCC-LI0001RT), the MHCC-97H (KG340) HCC cell line from Jiangsu KeyGEN BioTech Corp., Ltd (Nanjing, China), and the MHCC-97L HCC cell line gifted by the First Affiliated Hospital of Xi'an Jiaotong University. The human normal hepatocyte cell line L-02 was obtained from the Shanghai Institute of Cell Biology at the Chinese Academy of Sciences (Shanghai, China) and cultured as previously described [19 (link)]. The cell lines were authenticated using STR profiling and tested for mycoplasma contamination.
6
HCC Tissue Collection and Cell Culture
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A total of 90 paired tissues of HCC tumours and adjacent normal samples and related clinical information were obtained from patients undergoing liver resection at the Liver Transplantation Center in The First Affiliate Hospital of Nanjing Medical University between October 2012 and March 2013. This research was approved by our Institutional Ethics Committee. All patients in our study offered their informed consent to take part in our study prior to surgery. Fresh tissues were collected and immediately frozen in liquid nitrogen. The diagnosis of all patients was histopathologically confirmed. The human hepatoma cell lines, HepG2, SMMC‐7721, SNU‐423, Hep3B, Huh7, MHCC‐97H and the human normal L02 cell line were obtained from KeyGen (Nanjing KeyGen Biotech Co., Ltd., Jiangsu, China). The cells were cultured in DMEM medium (Gibco, Carlsbad, CA, USA) supplemented with 10% foetal bovine serum (FBS) and maintained in an atmosphere of 5% CO2 at 37°C.
7
Hepatocellular Carcinoma Tissue Collection
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116 paired pathologically diagnosed HCC samples (including tumor tissues and matched non-tumor liver tissues) were obtained between May 2010 and October 2014 at The Affiliated Changzhou NO.2 People's Hospital of Nanjing Medical University (Changzhou, Jiangsu, China). Written consent approving the usage of tissues in our study were obtained from each patient. The histopathological diagnoses of HCC were conducted by three experienced pathologists independently. This study was approved by the ethnic committee of the Affiliated Changzhou NO.2 People's Hospital of Nanjing Medical University ([2017]KY013-01). Human HCC cell lines (SMMC-7721, Huh7, HepG2, MHCC-97H, MHCC-97L, Hep3B) and the human normal liver L02 cell line were obtained from KeyGen (Nanjing KeyGen Biotech Co., Ltd., Jiangsu, China). All cells used in our study were cultured in Dulbecco's modified Eagle's medium (DMEM, Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA) and 1% streptomycin/penicillin at 37°C in a humidified environment consisting of 95% air and 5% CO2.
8
Silencing of RRM1, UCK2, and G6PD in Hepatoma Cells
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HepG2, Hep3B, MHCC‐97H, Huh‐7, and HCCLM3 human hepatoma and normal L02 cell lines were obtained from Nanjing KeyGen Biotech Co. Ltd. All cells were cultured with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM, Invitrogen Life Technologies), supplemented with 100 U/ml penicillin, 100 μg/ml streptomycin and 10% foetal bovine serum. RRM1 siRNA, UCK2 siRNA, G6PD siRNA or their respective negative controls were synthesized by Riobio, and sequences of siRNAs were listed below:
| siRNA | sequence |
|---|---|
| si‐RRM1#1 | 5′‐UCUUAAUCGCGUAUAAGGCTT‐3′ |
| si‐RRM1#2 | 5′‐CCCAACAGGAGGACAGCUUTT‐3′ |
| si‐UCK2#1 | 5′‐GCGGCGAGCCCUUCCUUAUTT‐3′ |
| si‐UCK2#2 | 5′‐GCCAGAAGCAGGUGGUCAUTT‐3′ |
| si‐G6PD#1 | 5′‐GCUCUACGAAGAUCUGGAATT‐3′ |
| si‐G6PD#2 | 5′‐GCAUUGCACAUCAACGGAUTT‐3′ |
9
Establishment and Maintenance of Liver Cancer Cell Lines
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HCC cell lines (MHCC-97L, MHCC-97H, Hep3B) and immortalized liver cell line (LO2) were procured from KeyGEN BioTECH (JiangSu, China). All cells were maintained in 10% fetal bovine serum (FBS, Gibco Life Technologies, Grand Island, NY, USA) supplemented with high glucose Dulbecco's modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA, USA) with the addition of 100 U/mL penicillin/streptomycin (Invitrogen, Carlsbad, CA, USA). All cultures were maintained in humidified environments of 5% CO2 at 37°C.
All animal experimental procedures were reviewed and approved by the Southern Medical University Animal Use and Care Committee (Guangdong, China). Female immunodeficient BALB/c-nu/nu mice weighing 20±2 g between 6 to 8 weeks of age were obtained from the Southern Medical University Medical Laboratory Animal Center and reared under specific-pathogen-free (SPF) conditions under a 12 h light/dark cycle. Food and water were provided ad libitum.
All animal experimental procedures were reviewed and approved by the Southern Medical University Animal Use and Care Committee (Guangdong, China). Female immunodeficient BALB/c-nu/nu mice weighing 20±2 g between 6 to 8 weeks of age were obtained from the Southern Medical University Medical Laboratory Animal Center and reared under specific-pathogen-free (SPF) conditions under a 12 h light/dark cycle. Food and water were provided ad libitum.
10
Comparative Study of HCC Cell Lines
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Human HCC cell lines including HepG2, Huh7, SMMC-772, MHCC-97H and Hep3B, the human normal liver cell line L02 and HEK293T cell line were acquired from KeyGen (Nanjing, Jiangsu, China). In this study, all cells were cultured in DMEM, which was supplemented with 10% FBS (Gibco), and were tested for mycoplasma contamination.
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