T-DXd was provided by Daiichi Sankyo Co., Ltd. (Tokyo, Japan). Other reagents used in this study were acquired from the indicated suppliers: trastuzumab (Chugai Pharmaceutical Corporation, Tokyo, Japan); KU-55933, VE-821, and UCN-01 (Merck Sigma-Aldrich, St. Louis, MO, USA); IFN-γ (R&D systems, Minneapolis, MN, USA); irinotecan (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan); ultra-LEAF purified human IgG1 isotype control recombinant antibody (BioLegend, San Diego, CA, USA); FITC mouse anti-human HER-2/neu (340553), FITC mouse IgG1 κ isotype control (555909), and 7-AAD (559925) (BD Biosciences, San Jose, CA, USA); PE anti-human HLA-ABC monoclonal antibody (W6/32) (12-9983-42), Alexa Fluor 488 anti-human CD326 (EpCAM) (53-8326-41), and PE mouse IgG2a κ isotype control (eBM2a) (12-4724-81) (Thermo Fisher Scientific, Waltham, MA, USA); HER2/ErbB2 rabbit mAb (#4290), Phospho-Akt rabbit mAb(#4060), Akt (pan) rabbit mAb (#4691), Phospho-ERK1/2 rabbit mAb (#4370), and ERK1/2 rabbit mAb (#4695) (Cell Signaling Technology, Danvers, MA, USA); anti-human HLA class I (HLA-ABC) mAb (D367-3) (Medical & Biological Laboratory, Aichi, Japan); Anti-β-actin antibody (sc-69879) (Santa Cruz Biotechnology, Dallas, TX, USA).
Irinotecan
Manufactured by Fujifilm
Sourced in Japan
Irinotecan is a chemical compound used in laboratory settings. It is a topoisomerase I inhibitor, which means it interferes with the action of the enzyme topoisomerase I, an enzyme involved in DNA replication and transcription. Irinotecan is commonly used in research applications to study the effects of topoisomerase I inhibition on cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Ku55933, by Merck Group (2 mentions)
Ve 821, by Merck Group (1 mentions)
Ucn 01, by Merck Group (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
7 aad 559925, by BD (1 mentions)
Phospho akt rabbit mab, by Cell Signaling Technology (1 mentions)
Akt pan rabbit mab, by Cell Signaling Technology (1 mentions)
Phospho erk1 2 rabbit mab, by Cell Signaling Technology (1 mentions)
Erk1 2 rabbit mab, by Cell Signaling Technology (1 mentions)
T dxd, by Daiichi Sankyo (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
Cisplatin, by Merck Group (1 mentions)
5 fluorouracil, by Merck Group (1 mentions)
Merlin, by Cell Signaling Technology (1 mentions)
Phospho histone h3, by Cell Signaling Technology (1 mentions)
Sunitinib, by Merck Group (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Doxorubicin, by Fujifilm (1 mentions)
4 protocols using irinotecan
1
Investigating T-DXd-Mediated Cytotoxicity in HER2+ Cells
2
Anticancer Compound Stock Preparation
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Hydroxyurea was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan) and dissolved in distilled water to prepare a 1 M stock solution. Gemcitabine, irinotecan, carboplatin and doxorubicin were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan) and dissolved in dimethylsulfoxide (DMSO) to prepare 1 mM, 20 mM, 25 mM and 10 mM stock solutions, respectively. Methotrexate was also purchased from Wako and dissolved in 1 M NaOH to prepare a 10 mM stock solution. Sunitinib, 5-fluorouracil, paclitaxel and cisplatin were purchased from Sigma (St. Louis, MO, USA) and dissolved in DMSO to prepare 10 mM, 10 mM, 1 mM and 100 mM stock solutions, respectively. Temozolomide was purchased from LKT Laboratories, Inc. (St. Paul, MN, USA) and dissolved in DMSO to prepare a 50 mM stock solution. Antibodies such as Cleaved Caspase-3 (Asp175, #9661), Cleaved PARP (Asp214, #9541), Merlin (#12888), Vimentin (#5741), phospho-Histone H3 (S10, #9706), Cleaved PARP (Asp214, Fluorescein conjugate, #9547), and GAPDH (#5174) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA).
3
Organoid Culture of Intestinal Tumors and Epithelia
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We performed organoid culture of intestinal tumors of Min mice and normal epithelia from wild‐type C57BL/6 mice. In brief, isolated tumor cells and epithelial cells were embedded in Matrigel on ice (growth factor‐reduced, phenol red‐free; BD Biosciences, San Jose, CA, USA) and seeded in 48‐well plates. The Matrigel was polymerized for 10 min at 37°C, and overlaid with 250 μL/well basal culture medium (advanced DMEM/F12 supplemented with penicillin/streptomycin, 10 mmol/L HEPES, Glutamax, 1 × N2, 1 × B27 [all from Thermo Fisher Scientific, Waltham, MA, USA] and 1 mmol/L N‐acetylcysteine [Sigma‐Aldrich, St. Louis, MO, USA]) containing the following optimized growth factor combinations: EGF and noggin for intestinal tumors; and EGF, noggin and Rspo1 for intestinal epithelia. For drug treatment, cells were seeded 2 days before drug administration and treated with 10 nM irinotecan (Wako, Pure Chemical Industries, Osaka, Japan) for 48 h. All experiments and procedures were approved by the Keio University Animal Research Committee, and all methods were carried out in accordance with the approved guidelines.
4
Assay of Prostate Cancer Cell Lines
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C4-2 cells, LNCaP cells, PC3 cells, and DU145 cells were purchased from the American Type Culture Collection and maintained at 37°C with 5% CO2 in RPMI (Wako) supplemented with 10% fetal bovine serum (FBS), 20 U/ml penicillin, and 100 µg/ml streptomycin. HEK293T cells were maintained at 37°C with 5% CO2 in DMEM (Wako) supplemented with 10% FBS, 20 U/ml penicillin, and 100 µg/ml streptomycin. PC3 cells and DU145 cells that stably express AR-FLAG were established by the selection with 50 µg/ml blasticidin S (Wako) after infection of lentivirus carrying AR-FLAG gene. Cells were treated with 1–10 µM irinotecan (Wako) at 37°C for 24 h, 10–50 µM etoposide (Sigma) at 37°C for 24 h, 100–1000 µM hydroxyurea (Abcam) at 37°C for 24 h, 10 µM Ku55933 (Sigma) at 37°C for 24 h, 1–10 µM enzalutamide (ChemScene) at 37°C for 48 h, and 100 µM mirin (Sigma) at 37°C for 4 h. Cells were stimulated with 100 ng/ml EGF (R&D systems) in 2% FBS for 24–48 h. For the cycloheximide chase assay, cells were treated with 25 µg/ml cycloheximide (Sigma) at 37°C.
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