Kyse450

Manufactured by American Type Culture Collection

Sourced in United States

The KYSE450 is a cell line derived from human esophageal squamous cell carcinoma. It is a laboratory tool used for research purposes.

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34 protocols using kyse450

Culturing Human ESCC Cell Lines

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Four human ESCC cell lines (Eca109, EC9706, KYSE150, KYSE450) and human normal esophageal epithelial cell line (Het-1A) were all procured from the ATCC (Manassas, VA, USA) and propagated at 37 °C in a humidified incubator of 5% CO₂. Cell lines were cultured in RPMI-1640 medium (Gibco, Carlsbad, CA, USA) with 10% FBS (Gibco) and 1% Pen/Strep mixture.

Xu S., Li X., Li L., Wang Y., Geng C., Guo F., Zhang T., Du A., Lu Z., Hui H, & Wang Q. (2021). CTCF-silenced miR-137 contributes to EMT and radioresistance in esophageal squamous cell carcinoma. Cancer Cell International, 21, 155.

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Pyrotinib and X-ray Radiation in Esophageal Cancer

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Human esophageal cancer cell lines TE-1, TE-10, KYSE30, EC109, KYSE150, and KYSE450 (ATCC, Manassas, VA, U.S.A.) were cultured in DMEM complete medium containing 10%FBS (Invitrogen, U.S.A.) in 5% CO₂ at 37°C. For Pyrotinib treatment, cells were washed with PBS and incubated with FBS-free medium containing Pyrotinib (Henrui Medicine, China) at indicated dose for 24 h. For X-ray radiation, cells were subjected to irradiation at a dose of 200 cGy/min by a 6 MV linear accelerator (Elekta, Sweded). If cells were subjected to both pyrotinib and irradiation, cells were treated with pyrotinib for 24 h (3 μg/ml for TE-1, 4 μg/ml for KYSE30 cells) followed by irradiation at indicated dose. The dosage for pyrotinib was determined by cell viability assay that showed that pyrotinib produced a cytotoxic effect on TE-1 cells with IC50 = 3.32 (Supplementary Figure S1A,B) and on KYSE30 cells with IC50 = 4.294 (Supplementary Figure S1C,D).

Lian X., Zhu C., Lin H., Gao Z., Li G., Zhang N., Cao B, & Kang Y. (2020). Radiosensitization of HER2-positive esophageal cancer cells by pyrotinib. Bioscience Reports, 40(2), BSR20194167.

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ESCC Cell Lines Maintenance and Depletion

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The 4 types of human ESCC cell lines, KYSE150, KYSE30, KYSE180, and KYSE450, were all bought from ATCC, and maintained in Dulbecco Modified Eagle Medium, supplemented with 10% of fetal bovine serum (FBS) in a 5% CO₂ incubator at 37°C.
The shRNAs were transfected into ESCC cells by lipofectamine 2000 (11668019; Invitrogen). Stable depleted ESCC cells were screened by lentivirus infection and used in animal assays.

Ren C., Zhou Z., Wang X., Hua X., Zou M, & Zhang X. (2020). SHCBP1 Promotes the Progression of Esophageal Squamous Cell Carcinoma Via the TGFβ Pathway. Applied Immunohistochemistry & Molecular Morphology, 29(2), 136-143.

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Enrichment and Treatment of ESCC Cells

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Human ESCC cell lines including EC9706, EC109, KYSE410, KYSE150, and KYSE450 were all available from the ATCC (Manassas, VA, USA) for cell culture with 5% CO₂ at 37 °C. RPMI‐1640 medium (Gibco, Carlsbad, CA, USA) was acquired commercially, with 1% Pen/Strep mixture and 10% FBS as supplements. Medium was changed every 3 days. After cells had reached about 80% confluence at the 3rd passage, CD133⁺ cancer cells were obtained by treating with MACS CD133 kit (Miltenyi Biotec, Teterow, Germany). ESCC cells without treatment were termed CD133^‐ cancer cells as control. About 2 mg·mL⁻¹ of actinomycin D was procured from Sigma‐Aldrich (St. Louis, MO, USA) to treat cells.

Wu D., He X., Wang W., Hu X., Wang K, & Wang M. (2020). Long noncoding RNA SNHG12 induces proliferation, migration, epithelial–mesenchymal transition, and stemness of esophageal squamous cell carcinoma cells via post‐transcriptional regulation of BMI1 and CTNNB1. Molecular Oncology, 14(9), 2332-2351.

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Esophageal Squamous Cell Carcinoma Cell Lines

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ESCC cell lines (KYSE150, KYSE270, KYSE450, TE4, and TE8) and a normal esophageal epithelial cell line (HET-1A) were obtained from ATCC (MA, USA). The Roswell Park Memorial Institute (RPMI)-1640 medium, 5-diphenyltetrazolium bromide (MTT) kit, and 4′,6-Diamidino-2-Phenylindole (DAPI) stain solution were supplied by Sigma (MO, USA). Lipofectamine 3000 reagents and Bicinchoninic Acid (BCA) kit were purchased from Thermo Scientific (CA, USA). Fetal Bovine Serum (FBS) was procured from GIBCO (CA, USA). Horseradish Peroxidase (HRP)-conjugated goat anti-rabbit/mouse antibodies were obtained from Cell Signaling Technology (MA, USA). Alexa Fluor 488 or 555 donkey anti-rabbit/mouse secondary antibodies were purchased from Beyotime (Shanghai, China). Information on primary antibodies used in this study is listed in Table S1.

Han G., Zhou S., Shen J., Yang Y., Bian X., Li Y., Ling R., Liang R, & Tao M. (2023). The role of TMEM26 in disrupting tight junctions and activating NF-κB signaling to promote epithelial-mesenchymal transition in esophageal squamous cell carcinoma. Clinics, 78, 100276.

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Induction of Autophagy and Ferroptosis in Human SCCs

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Human SCCs cell lines (KYSE410 and KYSE450) were purchased from ATCC (Manassas, VA, USA). Cells were maintained in RPMI-1640 (#PM150120; Procell, Wuhan, China) plus 10% fetal bovine serum (FBS; #SH30084.03; Hyclone, South Logan, UT, USA), 100 units/mL penicillin, 100 μg/mL streptomycin. Rapamycin (#ab120224; Abcam, Cambridge, MA, USA), Erastin (#ab209693; Abcam, Cambridge, MA, USA) and Gefitinib (Iressa, AstraZeneca, Macclesfield, UK) were dissolved in dimethyl sulfoxide (DMSO; Sigma, St. Louis, MO, USA) as well as stored at -20°C. To activate autophagy or ferroptosis, cells were administrated with 0.1 μM Rapamycin or Erastin for 16 h.

Chen L., Niu X., Qiao X., Liu S., Ma H., Shi X., He X, & Zhong M. (2022). Characterization of Interplay Between Autophagy and Ferroptosis and Their Synergistical Roles on Manipulating Immunological Tumor Microenvironment in Squamous Cell Carcinomas. Frontiers in Immunology, 12, 739039.

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Esophageal cancer cell lines treated

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Human ESCC cells, KYSE510, KYSE450, and KYSE70, were obtained from ATCC (Manassas, VA, USA) and ECACC (Porton Down, Salisbury, UK) with proper authentication. KYSE510 and KYSE450 cells were exposed to ethanol. KYSE70 cells were treated with 5-aza-2’-deoxycytidine (DAC; Sigma-Aldrich, St. Louis, MO, USA) for 72 h.

Xiong Z., Ren S., Chen H., Liu Y., Huang C., Zhang Y.L., Odera J.O., Chen T., Kist R., Peters H., Garman K., Sun Z, & Chen X. (2017). PAX9 regulates squamous cell differentiation and carcinogenesis in the oro-esophageal epithelium. The Journal of pathology, 244(2), 164-175.

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Esophageal Cancer Cell Lines Characterization

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Esophageal cancer cell lines KYSE‐150, KYSE‐450, and ECA109 and normal esophageal HEEC cell lines were purchased from ATCC company.

Liu X., Wu W., Zhang S., Tan W., Qiu Y., Liao K, & Yang K. (2021). Effect of miR‐630 expression on esophageal cancer cell invasion and migration. Journal of Clinical Laboratory Analysis, 35(6), e23815.

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Esophageal Squamous Cell Carcinoma Cell Lines

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Establishing Esophageal Cancer Cell Lines

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Six human ESCC cell lines—TE1, ECA109, EC9706, KYSE30, KYSE150 and KYSE450—were purchased from American Type Culture Collection (Manassas, VA, USA). Based on their characteristics and a scholarly literature review, we selected these six cell lines for use in our current study to minimize the known differences that influence radiation response. All of these cell lines have been established in countries marked by a higher incidence of esophageal cancer and were histologically derived from squamous cell carcinomas. Additionally, none of the cell lines that were donated had been exposed to X-ray irradiation prior to surgery [15 (link), 16 ]. All of these cells lines were maintained in their recommended growth medium and incubated in a humidified 5% CO₂ atmosphere at 37°C. Each cell line was regularly observed and tested to avoid possible mycoplasma contamination.

Jin Y.Y., Chen Q.J., Wei Y., Wang Y.L., Wang Z.W., Xu K., He Y, & Ma H.B. (2016). Upregulation of microRNA-98 increases radiosensitivity in esophageal squamous cell carcinoma. Journal of Radiation Research, 57(5), 468-476.

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