Sk hep1

Manufactured by American Type Culture Collection

Sourced in United States, China, Japan, Australia

SK-Hep1 is a human hepatocellular carcinoma cell line. It is a commonly used in vitro model for liver cancer research.

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SK‐hep1 cell line (2 citations)

378 protocols using sk hep1

Establishment of AR knockdown cell lines

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The human HCC cells were maintained in DMEM (Invitrogen, Grand Island, NY) with 10% fetal bovine serum (FBS), 1% Glutamine, and 1% penicillin/streptomycin. NK-92MI cells (ATCC, Manassas, VA) were maintained in α-MEM (Invitrogen) with 0.2 mM inositol, 0.1 mM 2-mercaptoethanol, 0.02 mM folic acid, horse serum to a final concentration of 12.5% and FBS to a final concentration of 12.5% based on ATCC guidelines. All cell lines were cultured ina5%(v/v) CO2 humidified incubator at 37 °C The SK-Hep1 (ATCC, Manassas, VA) and SNU423 (ATCC, Manassas, VA) AR stable transfectants were established based on a previous procedure [14 (link)]. Cisplatin (479306), MG132 (M8699) and Cycloheximide (CHX, 227048) were purchased from Sigma.
To generate AR knock-down stable clones of SK-Hep1 (ATCC, Manassas, VA) and SNU423 cells (ATCC, Manassas, VA), HEK-293T cells were transfected with lentiviral vectors, pLKO1-sh-AR/pLKO1-scr, with the psAX2 packaging plasmid, and pMD2G envelope plasmid for 48 hrs to obtain the lentivirus supernatant, which was frozen at −80 °C for later use.
For the luciferase reporter assay, cells were transfected using Lipofectamine 3000 (Invitrogen) reverse transfection protocol, according to the manufacturer's instructions. See Supplemental data for detailed sequence information.

Shi L., Lin H., Li G., Sun Y., Shen J., Xu J., Yeh S., Cai X, & Chang C. (2016). Cisplatin enhances NK cells immunotherapy efficacy to suppress HCC progression via altering the androgen receptor (AR)-ULBP2 signals. Cancer letters, 373(1), 45-56.

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Characterization of HCC Cell Lines

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HCC cell Huh-7 and BEL-7404 were purchased from the ATCC. Hepatoblastoma cell Sk-hep-1 and endothelial-derived cell Sk-Hep1 were purchased from the ATCC. All the above cell lines of HCC were cultured in DMEM (Biological Industries) þ 10% FBS (Biological Industries) þ 1% penicillin/streptomycin at 37 C with 5% CO 2 and saturated humidity. All cell lines used were cultured within 35 generations and regularly tested for Mycoplasma contamination by the Plasmo Test kit (InvivoGen, rep-pt1). The short tandem repeat analysis method was used to verify the identity of cell lines twice a year at the core institution.

Du L., Li Y., Kang M., Feng M., Ren Y., Dai H., Wang Y., Wang Y, & Tang B. (2021). USP48 Is Upregulated by Mettl14 to Attenuate Hepatocellular Carcinoma via Regulating SIRT6 Stabilization. Cancer research, 81(14).

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Colchicine Derivative TCD Evaluation

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TCD (C₃₀H₂₇NO₇S), an odorless, yellow crystal powder, is a novel colchicine derivative; it was purchased from PUMC Pharmaceutical Co., Ltd. (Bejing, China) for this study. This water-insoluble compound was dissolved in DMSO (Sigma-Aldrich, St. Louis, MO, USA) to be used as a stock solution for further in vitro studies. Three human hepatoma cell lines, Hep-J5, Mahlavu, and SK-Hep-1, were kindly provided by Professor Kwang-Huei Lin [26 (link), 27 ], among which SK-Hep-1 was originally purchased from the American Type Culture Collection, VA, USA [26 (link)], and J5 and Mahlavu were originally obtained from Dr. C.S Yang [28 (link)], National Taiwan University, and Dr. C. P. Hu [27 ], Veterans General Hospital, Taiwan, respectively. Mahlavu was originally established from a female hepatoma patient in 1972 and has been in continuous culture since then [29 (link)]. Huh7 was purchased from the Japanese Collection of Research Bioresources Cell Bank, Osaka, Japan. Hep-G2 and Hep-3B were purchased from the Bioresource Collection and Research Center, Hsinchu, Taiwan. Dulbecco’s modified Eagle medium (DMEM) and fetal bovine serum (FBS) were obtained from Sigma-Aldrich and Biological Industries (South Logan, UT, USA), respectively. All other chemicals were purchased from Sigma-Aldrich.

Ho C.T., Chang Y.J., Yang L.X., Wei P.L., Liu T.Z, & Liu J.J. (2015). A Novel Microtubule-Disrupting Agent Induces Endoplasmic Reticular Stress-Mediated Cell Death in Human Hepatocellular Carcinoma Cells. PLoS ONE, 10(9), e0136340.

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Cultivation of Human Hepatocellular Carcinoma Cell Lines

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The human HCC cell lines (Sk-hep1, Huh7, L02, Hep-G2, LM3, HA22T, JHH7) were provided by the American Tissue Culture Collection (Manassas, Virginia, USA). All cell lines were maintained in Dulbecco's modi ed Eagle's medium (DMEM) (Gibco) and supplemented with 10% fetal bovine serum (FBS) (Gibco), 10 U/mL penicillin and 10 mg/mL streptomycin. Cells were grown in a humidi ed atmosphere incubator at 37°C in 5% carbon dioxide. All cells were cultured according to the manufacturer's guidelines.

Maher H., Pan Y., Kwabena G.O., Zhang B., Wang Y., Lv J., & Cai X. (2021). A Novelty of Long Non-coding RNA LOXL1-AS1: Suppresses Tumor Progression and Metastasis and an Independent Favorable Prognostic Factor in Hepatocellular Carcinoma.

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Cell Culture and Delipidized Medium

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MDA-MB-231, HeLa, HCT116, HCT-8, HCC1806, SKHEP-1, HAEC, HPAC, HepG2, Huh7, and 293T were obtained from the American Tissue Culture Collection. HCCLM3, L02, and KYSE150 were obtained from China Center for Type Culture Collection. HCT-8, HCC1806, and KYSE150 cells were cultured in 1640 (Corning), and other cells were cultured in DMEM (Corning). The above mediums were supplemented with 10% FBS (#900-108; Gemini) and 100 U/ml penicillin and streptomycin (#15140122; Gibco). All cells were cultured in a humidified atmosphere of 5% CO₂ at 37°C.
For delipidized medium, cells were first seeded in the medium containing regular 10% FBS and were switched into the medium containing 10% delipidized FBS (#P30-3402; PAN) upon treatment on the following day.

Chu Q., An J., Liu P., Song Y., Zhai X., Yang R., Niu J., Yang C, & Li B. (2022). Repurposing a tricyclic antidepressant in tumor and metabolism disease treatment through fatty acid uptake inhibition. The Journal of Experimental Medicine, 220(3), e20221316.

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Culturing Liver Cancer Cell Lines

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Liver cancer cell lines (SK-Hep-1, Huh7, HepG2, HCCLM3) were purchased from the American Tissue Culture Collection (Manassas, VA, USA) and cultured in accordance with the recommended guidelines. Sorafenib-resistant HCC cell lines were cultured with Sorafenib as previously reported (40 (link), 41 (link)).

Pan Y., Hu G.Y., Jiang S., Xia S.J., Maher H., Lin Z.J., Mao Q.J., Zhao J., Cai L.X., Xu Y.H., Xu J.J, & Cai X.J. (2021). Development of an Aerobic Glycolysis Index for Predicting the Sorafenib Sensitivity and Prognosis of Hepatocellular Carcinoma. Frontiers in Oncology, 11, 637971.

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Culturing Cancer Cell Lines

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The human gastric cancer cell lines, SNU-16, SNU-484, and SNU-668, were purchased from Korean Cell Line Bank (KCLB, Seoul, Korea). The human liver cancer cell lines, SK-Hep-1, HepG2, and Hep3B, were obtained from American Tissue Culture Collection (ATCC, Manassas, VA). All of the cancer cell lines were adapted to RPMI 1640 (WelGENE, Daegu, Korea) containing 10% fetal bovine serum (FBS) (WelGENE) and maintained as monolayer cultures at 37℃ in a humidified, 5% CO₂ environment.

Park J.W., Baek N.S., Lee S.C., Oh S.J., Jang S.H., Kim I.H., Kim D.Y, & Kim H.K. (2014). Preclinical Efficacy Testing for Stomach and Liver Cancers. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 46(2), 186-193.

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Human Hepatocellular Carcinoma Cell Line Experiments

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We purchased human HCC cell lines (HepG2, SMMC7721, Huh-7, and Sk-Hep-1) and a normal human liver cell line (L02) from the American Tissue Culture Collection (Manassas, VA, USA). We cultured cells in Roswell Park Memorial Institute 1640 medium (HyClone, Logan, UT, USA) containing 10% fetal bovine serum (FBS; Gibco, Gaithersburg, MD, USA) in a humidified atmosphere with 5% CO₂ at 37°C. We transfected miR-548c-3p inhibitors, lentiviral-stabilized hsa_circ_0101145 silenced vector (si-circ0101145), miR-548c-3p mimics, LAMC2 overexpression vector (FOSL2), and the negative controls (NCs) into cultured Huh-7 and HepG2 cells prior to subsequent experiments. We purchased lentiviral-based short hairpin RNA (shRNA) targeting hsa_circ_0101145 and lentiviruses overexpressing LAMC2 from GeneChem (Shanghai, China).

Jin J., Liu H., Jin M., Li W., Xu H, & Wei F. (2020). Silencing of hsa_circ_0101145 reverses the epithelial-mesenchymal transition in hepatocellular carcinoma via regulation of the miR-548c-3p/LAMC2 axis. Aging (Albany NY), 12(12), 11623-11635.

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Culturing Human Liver Cell Lines

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Human normal liver cell line (L02) and the human HCC cell lines (SMMC7721, Sk-Hep-1, HepG2, Huh-7, and HCCLM3) were purchased from the American Tissue Culture Collection (ATCC, Manassas, VA, USA). They were cultured in Roswell Park Memorial Institute-1640 medium (RPMI-1640; HyClone, Logan, UT, USA) containing 10% fetal bovine serum (FBS, Gibco, USA) in a humidified atmosphere with 5% v/v CO₂ at 37°C.

Lin T., Dai Y., Guo X., Chen W., Zhao J., Cao L, & Wu Z. (2019). Silencing Of hsa_circ_0008450 Represses Hepatocellular Carcinoma Progression Through Regulation Of microRNA-214-3p/EZH2 Axis. Cancer Management and Research, 11, 9133-9143.

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Culturing Cancer Cell Lines

Cited 1 time

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The colorectal cancer cell line HCT-116, prostate cancer cell line PC-3, and hepatocellular carcinoma cell line SK-Hep-1 were purchased from the American Type Cell Culture Collection (Manassas, VA, USA). Cell culture was performed following the procedure of our previous reports [6 (link)]. In summary, the cells were maintained in DMEM medium containing fetal bovine serum (FBS), penicillin, and streptomycin in humidified air containing 5% CO₂ at 37 °C.

Safwan S., Wang S.W., Hsiao G., Hsiao S.W., Hsu S.J., Lee T.H, & Lee C.K. (2022). New Trichothecenes Isolated from the Marine Algicolous Fungus Trichoderma brevicompactum. Marine Drugs, 20(2), 80.

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