Hcclm3

Manufactured by Thermo Fisher Scientific

Sourced in United States, China

The HCCLM3 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed for use in scientific research and analysis applications. The core function of the HCCLM3 is to provide a controlled environment for sample preparation and testing.

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Lab products found in correlation

67 protocols using hcclm3

Molecular Profiling of Liver Hepatocellular Carcinoma

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Thirty-five LIHC specimens and matched paracancerous normal tissues were obtained from patients diagnosed with LIHC at our hospital. All patients provided informed consent, and the ethics committee of the hospital approved our study. Patient characteristics are shown in Table 1. LIHC cell lines (Hep3B, HCCLM3, Huh7, and SNU-182) and normal liver epithelial cells (THLE2) were provided by American Type Culture Collection (ATCC, USA). Hep3B, HCCLM3, and Huh7 cells were cultured in DMEM (Gibco, USA). SNU-182 and THLE2 cells were cultured in medium RPMI-1640 (Gibco). All cells were incubated with 10% FBS at 37 °C and 5% CO₂. siRNA targeting lncRNA ST8SIA6-AS1 or HMGA1 (si-lnc or si-HMGA1), miR-142-3p mimics/inhibitor (anti-miR), and their negative controls (NC) (RiboBio Co. Ltd., China) were used to treat HCCLM3 and Huh7 cells using Lipofectamine 2000 (Invitrogen, USA). After 48 h, further functional analyses were performed.Table 1

Baseline characteristics of 35 HCC patients.

Categories	Cases (Total n = 35)	Percentage (%)
Gender
Male	14	40.0
Female	21	60.0
Age (years)
≥ 55	18	51.4
< 55	17	48.6
Tumor size (cm)
≥ 5	11	31.4
< 5	24	68.6
TNM stage
I + II	20	57.1
III + IV	15	42.9
Lymph node metastasis
Positive	19	54.3
Negative	16	45.7
Liver cirrhosis
Presence	13	37.1
Absence	22	62.9
Serum AFP (ng/mL)
≤ 20	18	51.4
> 20	17	48.6

Feng T., Yao Y., Luo L., Zou H., Xiang G., Wei L., Yang Q., Shi Y., Huang X, & Lai C. (2023). ST8SIA6-AS1 contributes to hepatocellular carcinoma progression by targeting miR-142-3p/HMGA1 axis. Scientific Reports, 13, 650.

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Establishing TRAIL-resistant Liver Cancer Cell Lines

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Huh-7 cell line (3111C0001CCC000679) was obtained from China Center for Type Culture Collection (CCTCC; Beijing, China) and Dulbecco’s Modified Eagle’s Medium (DME-H-21 4.5 g/Liter Glucose; Invitrogen) supplemented with 10% FBS (Invitrogen). HCCLM3 (C6303) was obtained from Beyotime (Shanghai, China) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% FBS (Invitrogen).
The human liver cancer cell line Huh-7 and HCCLM3 were exposed to gradually increased concentrations of recombinant human TRAIL (rhTRAIL; 1, 10, 100, and 1,000 ng/ml) as described previously (26 (link), 33 (link), 34 (link)) for establishing the TRAIL-resistant Huh-7 (R) and HCCLM3 (R).

Sun J., Xu H., Lei Z., Li Z., Zhu H., Deng Z., Yu X., Jin X, & Yang Z. (2022). The lncRNA CASC2 Modulates Hepatocellular Carcinoma Cell Sensitivity and Resistance to TRAIL Through Apoptotic and Non-Apoptotic Signaling. Frontiers in Oncology, 11, 726622.

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Investigating miRNA-590-5P and OLR1 in HCC

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Human HCC cell lines [Hep3B (RRID: CVCL_XE51), HepG2 (RRID: CVCL_0027), Huh-7 (RRID: CVCL_0336), HCCLM3 (RRID: CVCL_6832), and SK-HEP-1 (RRID: CVCL_0525)] were purchased from the Chinese Academy of Medical Sciences (Beijing, China). Hep3B, HepG2, Huh-7, HCCLM3, and SK-HEP1 cells were cultured in RPMI 1640 Medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS (Invitrogen, Carlsbad, CA, USA) and 1% penicillin/streptomycin at 37 °C in 5% CO₂.
Small interference RNAs (si-RNAs) targeting miRNA-590-5P (si-miRNA-590-5P) and OLR1 (si-OLR1), scramble control (si-control), miRNA-590-5P mimic, and its scramble control (miR-control) were obtained from GenePharma Co. Ltd (Suzhou, China). All these plasmids were transfected into HepG2 or HCCLM3 cells by Lipofectamine^® 2000 reagent (Invitrogen, Carlsbad, CA, USA).

Zhou M., Zhang G., Hu J., Zhu Y., Lan H., Shen X., Lv Y, & Huang L. (2021). Rutin attenuates Sorafenib-induced Chemoresistance and Autophagy in Hepatocellular Carcinoma by regulating BANCR/miRNA-590-5P/OLR1 Axis. International Journal of Biological Sciences, 17(13), 3595-3607.

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Comparative Study of HepG2 and HCCLM3 Hepatoma Cell Lines

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HepG2 cell lines (HB-8065) was purchased from the American Type Culture Collection (ATCC). The malignant human hepatoma cell line HCCLM3 was obtained from Nanjing Kebai Biotechnology Co., Ltd (CBP60654). HepG2 and HCCLM3 cells lines were cultured in high-glucose DMEM (Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco), 100 U/mL penicillin, and 100 μg/mL streptomycin.

Wang S., Chen L, & Liu W. (2022). Matrix stiffness-dependent STEAP3 coordinated with PD-L2 identify tumor responding to sorafenib treatment in hepatocellular carcinoma. Cancer Cell International, 22, 318.

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Cell Line Culture Protocols for Liver Cancer Research

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HEK293T cells, Human HCC cell line Huh7, HCCLM3, JHH-7 and PLC/PRF/5 were purchased from the Chinese Academy of Sciences Cell Bank (Shanghai, China). HEK293T, Huh7, HCCLM3, JHH-7 were cultured with Dulbecco's modified Eagle's medium (DMEM) (Gibco) containing 10% (vol/vol) Fetal Bovine Serum (FBS) (ExCell Bio, FSP500) and 1% (vol/vol) Penicillin–Streptomycin (P/S) (KeyGen, KGY0023). PLC/PRF/5 were cultured with Minimum Essential Medium (MEM, Gibco) containing 10% (vol/vol) FBS, 1% (vol/vol) Non-Essential Amino Acids Solution (NEAA) (Gibco), 1% (vol/vol) L-Glutamine (Gibco) and 1% (vol/vol) P/S. All the cells were incubated in a humidified-atmosphere incubator of 5% CO₂ at 37 °C.

Tao S., Xie S.J., Diao L.T., Lv G., Hou Y.R., Hu Y.X., Xu W.Y., Du B, & Xiao Z.D. (2023). RNA-binding protein CCDC137 activates AKT signaling and promotes hepatocellular carcinoma through a novel non-canonical role of DGCR8 in mRNA localization. Journal of Experimental & Clinical Cancer Research : CR, 42, 194.

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Hepatocellular Carcinoma Patient-Derived Tissues

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A total of 40 HCC and paraneoplastic tissues were obtained from patient volunteers who had their tumors removed at the Wuhan University Central South Hospital and Qilu Hospital of Shandong University. All patients provided their written informed consent. All tumor samples were histopathologically verified as HCC by 2 senior independent pathologists. This study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of Wuhan University Zhongnan Hospital. HEK293T, Huh-7, HCC-LM3, Hep-3B, and Hep-G2 cells were purchased by the Stem Cell Bank, Chinese Academy of Sciences (Shanghai, China). All cell lines were verified by short tandem repeat profiling and routinely tested for mycoplasma contamination. All cells were cultured in a 37 °C, 5% CO₂ incubator. HEK293T, Huh-7, HCC-LM3, and Hep-G2 cells were cultured in the Dulbecco's modified Eagle medium (Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (BD Bioscience, San Jose, CA, USA). Hep-3B cells were cultured in the minimum essential medium (Gibco) supplemented with 10% fetal bovine serum, 1% GlutaMAX (Gibco), 1% nonessential amino acids (Gibco), and 100 mM solution of 1% sodium pyruvate (Gibco).

Zhang H., Zhai X., Liu Y., Xia Z., Xia T., Du G., Zhou H., Franziska Strohmer D., Bazhin A.V., Li Z., Wang X., Jin B, & Guo D. (2023). NOP2-mediated m5C Modification of c-Myc in an EIF3A-Dependent Manner to Reprogram Glucose Metabolism and Promote Hepatocellular Carcinoma Progression. Research, 6, 0184.

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Culturing Human Liver and Kidney Cells

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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).

Lai M., Li D., Liu M., Zhang R., Wang L., Peng W., Xu H., Wu S., Liang S., Gu Y., Nan A, & Zeng X. (2023). CircFAM114A2 inhibits the progression of hepatocellular carcinoma via miR‐630/HHIP axis. Cancer Medicine, 12(11), 12553-12568.

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HCC Cell Culture and Drug Intervention

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The HCCLM3 cells were purchased from the China Center for Type Culture Collection (CCTCC), and the Huh7 cells were purchased from Suyan Biotechnology Co., Ltd. The THLE-2 cells were purchased from Meisen Cell Technology Co., Ltd. The HCCLM3 and Huh7 Cells were cultured in DMEM/high-glucose medium (Gibco; Thermo Fisher Scientific, Inc.) with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) and 1% penicillin-streptomycin (Gibco; Thermo Fisher Scientific, Inc.) at 37°C and 5% CO₂. The THLE-2 cells require a special coating medium (CTCC-002-30, Meisen Cell Technology Co., Ltd.) and were incubated overnight in an incubator at 37°C and 5% CO₂ and cultured in BEGM™ BulletKit™ medium (cat. no. CC-3170, Lonza Group, Ltd.) with 10% FBS. The HCC cells were observed and representative images were captured under an inverted microscope (Ts2R-FL, Nikon Corporation; ×100 magnification) at 0 and 24 h following drug intervention. The HCC cells were treated with PPVI (4 µM) at 37°C for 24 h, following which different assays were performed. Inhibitors, such as 3-MA (1 mM), Ac-DEVD-CHO (50 µM) and Fer-1 (1 µM) for 2 h prior to treatment with PPVI (4 µM) for 24 h.

Huang Q., Li J., Ma M., Lv M., Hu R., Sun J., Zhong X., Sun X., Feng W., Ma W., Zhang W., Zhan B., Han Z, & Zhou X. (2023). High-throughput screening identification of a small-molecule compound that induces ferroptosis and attenuates the invasion and migration of hepatocellular carcinoma cells by targeting the STAT3/GPX4 axis. International Journal of Oncology, 62(3), 42.

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CCDC58 Knockdown in Liver Cancer Cells

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The human liver cancer cell line (MHCC97-H) was purchased from Zhong Qiao Xin Zhou Biotechnology Co., Ltd (Shanghai, China). Human liver cancer cell lines (Huh-7, HCCLM3, and Hep 3B) and human liver cell line (WRL-68) were purchased from Haixing Biological Technology Co., Ltd (Suzhou, China). The human liver cancer cell line (Hep G2) was purchased from Procell Life Science&Technology Co., Ltd (Wuhan, China). All cell lines were identified correctly by STR analysis. MHCC97-H, Huh-7, HCCLM3, and WRL-68 were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, USA) + 10%Fetal Bovine Serum (FBS, Gibco, USA) + 1%Antibiotics (Penicillin streptomycin, Gibco, USA). Hep G2 and Hep 3B were cultured in Minimum Essential Medium (MEM, Gibco, USA) + 10%Fetal Bovine Serum (FBS, Gibco, USA) + 1%Antibiotics (Penicillin streptomycin, Gibco, USA). The cell incubator was incubated at 37 °C with 5% CO₂. Cells in 6-well plates were transiently transfected with small-interfering RNA (siRNA) of CCDC58 and its negative control at 70–80% density using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer’s description. The siRNA sequences were presented in Table S1. After 48 h transfection, cells were tested for knockdown efficiency of CCDC58 and used for functional experiments.

Yang K., Ma Y., Chen W., Liu L., Yang Z., He C., Zheng N., Liu X., Cheng X., Song J., Chen Y., Qiao H, & Zhang R. (2024). CCDC58 is a potential biomarker for diagnosis, prognosis, immunity, and genomic heterogeneity in pan-cancer. Scientific Reports, 14, 8575.

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Cultivation of Human Liver Cancer Cell Lines

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Human HCC cell lines (HCCLM3, Hep-G2, Hep3B, HuCCT1, Huh-1, Huh-7, JHH-7, MHCC97-H, PLC/PRF/5, SNU-761, SNU-878) were obtained from different sources (ATCC: American Type Culture Collection; ZHFU: Zhongshan Hospital Fudan University; JCRB: Japanese Collection of Research Bioresources; SIBS: Shanghai Institutes of Biological Sciences; and KCLB: Korean Cell Line Bank). All the cells were cultured in ATCC recommended growth media (HCCLM3, Hep G2, Huh-1, Huh-7, MHCC97-H, and PLC/PRF/5 in DMEM containing 10% fetal bovine serum (FBS); Hep3B in MEM containing 10% FBS and 0.01mM NEAA; HuCCT1 in RPMI1640 containing 10% FBS; JHH-7 in William’s E containing 10% FBS; SNU-761 and SNU-878 in RPMI1640 containing 10% FBS), in 5% CO₂ at 37ºC. All culture media were obtained from GIBCO or Hyclone. HepG2 is HBV negative, but HCV positive; Huh-1, HCCLM3, and PLC/PRF/5 are HBV positive, and HCV negative.

Zhao Y., Lau L.F., Dai X, & Li B. (2016). In Vitro and In Vivo Anticancer Activity of Gimatecan against Hepatocellular Carcinoma. Asian Pacific Journal of Cancer Prevention : APJCP, 17(11), 4853-4856.

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