SW620 cells stably transfected with SOX2 short hairpin RNA (shRNA) or negative shRNA (5 × 106 cells in 0.1 mL PBS) were subcutaneously injected into the right dorsal flanks of 4-week-old female Balb/c nude mice (n = 3/group), respectively. Twenty-two days later, mice bearing tumors were administered intraperitoneal injection of PBS, Chloroquine (Sigma-Aldrich, C6628, CQ, 20 mg/kg), irinotecan (Sigma-Aldrich, I1406, 20 mg/kg), or combination of irinotecan and CQ every 3 days. Tumor size was measured with a digital caliper every 3 days. Tumor volume was calculated from digital caliper measurements of tumor dimensions in mm using the formula for a prolate ellipsoid: (L × W2)/2, where L is the longer of the 2 measurements. At the endpoint, tumors were harvested and weighed. The excised tissues were either fixed in 10% neutral-buffered formalin or snap frozen in liquid nitrogen. Tumor sections from paraffin-embedded blocks were used for histologic examination. All animal experiments were conducted as per the protocol approved by the Animal Care and Use Committee of the Southern Medical University.
Irinotecan
Manufactured by Merck Group
Sourced in United States, Germany, France
Irinotecan is a topoisomerase I inhibitor used in the laboratory setting for various research applications. It functions by interfering with the enzyme topoisomerase I, which is involved in DNA replication and transcription. Irinotecan can be used to study its effects on cellular processes and as a research tool in drug discovery and development.
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Lab products found in correlation
Oxaliplatin, by Merck Group (14 mentions)
Cisplatin, by Merck Group (13 mentions)
Etoposide, by Merck Group (9 mentions)
5 fluorouracil, by Merck Group (8 mentions)
Gemcitabine, by Merck Group (8 mentions)
Sn 38, by Merck Group (7 mentions)
Doxorubicin, by Merck Group (7 mentions)
5 fluorouracil 5 fu, by Merck Group (6 mentions)
Paclitaxel, by Merck Group (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Topotecan, by Merck Group (4 mentions)
Cyclophosphamide, by Merck Group (4 mentions)
Ht 29, by American Type Culture Collection (4 mentions)
Hct116, by American Type Culture Collection (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Merck Group (3 mentions)
Vinorelbine, by Merck Group (3 mentions)
Vincristine, by Merck Group (3 mentions)
Staurosporine, by Merck Group (3 mentions)
Docetaxel, by Merck Group (3 mentions)
79 protocols using irinotecan
1
Evaluating SOX2 Knockdown and Chemoresponse in Xenograft Mice
2
Angiogenesis Inhibitors and Chemotherapeutics
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Recombinant human VEGF 165 was obtained from PromCell (Rocky Hill, NJ, USA). Sunitinib, temsirolimus, interferon 2αΑ, irinotecan, SN38 (the active metabolite of irinotecan), poly-2-hydroxyethyl methacrylate (polyHEMA), sodium orthovanadate, and gelatin were purchased from Sigma-Aldrich (St Quentin Fallavier, France). CellTracker Green, collagen, Enhanced chemiluminescence detection reagents, the micro–bicinchoninic acid (micro-BCA) protein assay reagent kit were purchased from ThermoFisher Scientific (Courtaboeuf, France). Nitrocellulose was obtained from Schleicher & Schuell BioScience (Perkin-Elmer Life Science, FRANCE).
3
Organoid-Based Chemo-Sensitivity Screening
4
Irinotecan Toxicity in Animal Model
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Animals of this group were administered a single IP injection of 200 mg/kg of irinotecan in a sorbitol/lactic acid buffer.14 irinotecan was purchased from Sigma Aldrich Chemical Co (Cat # 1347609 USP Sigma Aldrich). The animals were then kept under standard housing conditions for 4 weeks without further treatment.
5
Establishment of Drug-Resistant C6 Glioma Cells
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The rat C6 glioma cell line was maintained at 37˚C/5% CO2 in Dul e o's Modified Eagle Medium (DMEM) (Invitrogen) with 10% foetal bovine serum (FBS). Drug selected cell lines (C6-etoposide and C6-irinotecan) were established by continuous culture over 5 passages at the drug concentration established to cause death of 70% of cells i.e.in the presence of 16 M etoposide "ig a a d 10 M irinotecan (Sigma). In the case of etoposide selection, cells were incubated for 2 hours [15] and then media was changed while for irinotecan selection, cells were incubated for 96 hours then media was changed [16] . Similarly, in experiments with prolonged duration such as clonogenic assays, the drugs were removed after 2 hours and 96 hours for etoposide and irinotecan, respectively.
6
Monoclonal Antibody for EGFR Detection
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The rat monoclonal antibody ICR62 (IgG2b) was raised against the external domain of the EGFR on the breast carcinoma cell line (MDA-MB468) [61 (link)]. The secondary antibody used in this study included FITC-conjugated rabbit anti-mouse IgG Star 9B (Bio-rad Antibodies, Kidlington, UK). The cytotoxic drugs, 5-FU, Irinotecan and Oxaliplatin were purchased from Merck, UK.
7
Comparative Analysis of Hepatic Cell Lines and Natural Product Compounds
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The hepatic cancer cell lines, Hep3b and HepG2 and the normal human fibroblast cell line CRL1554 were obtained from the American Type Culture Collection (Manassas, VA, USA). HepG2 and Hep3b cells were grown in Eagle's minimal essential medium (EMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.) in a 5% CO2 incubator at 37°C. HepG2 cells, which were originally thought to be a HCC cell line (12 (link),13 (link)), have been misidentified and are considered to be a hepatoblastoma cell line (14 (link)). CRL1554 cells were grown in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc.) containing 10% FBS in a CO2 incubator at 37°C. Penicillin/streptomycin (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), gentamicin (Gibco; Thermo Fisher Scientific, Inc.), L-glutamine (Fluka; Honeywell Labs, Muskegon, MI, USA) and sodium hydrogen carbonate were added to all media during preparation. The NPCs were all obtained from Sigma-Aldrich (Merck KGaA) and included homoharringtonine (HHG), Kmf, Cur, Que, Rsv, hesperetin (Hsp), betulinic acid (BetA), indol-3-carbinol (IC3), coumarin (Cmr), sulanidac (Sul), irinotecan (Irt), lycopene (Lyp), silibinin (Sil) and sinirgin (Snn).
8
Chemosensitivity Screening of Colorectal Cancer Organoids
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LMT organoids in good condition were harvested, passaged, and seeded in 96-well cell culture plates. The organoid density was adjusted to 50–60/10 μL Matrigel with 200 μL culture medium. For drug testing, the organoid culture medium was removed and replaced with a 200 μL drug-containing culture medium: 2.5 mg/mL cetuximab (Erbitux, Merck), irinotecan (I1406, Merck), or oxaliplatin (O9512, Sigma). Organoids were photographed seven days after drug treatment (EVOS FL Cell Imaging System, Thermo Fisher Scientific), and cell viability was also evaluated at seven days by the CellTiter 96 Aqueous One Solution cell assay (Promega, G3580) according to the manufacturer's instructions.
9
Modulation of Cellular Pathways by Bile Acids
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LCA (cat # L6250; Sigma-Aldrich, St. Louis, MI, USA) was dissolved in DMSO at a stock concentration of 100 mM. LCA was used at a concentration of 0.03 µM, corresponding to normal human serum concentration [7 (link), 47 , 48 (link)]. Non-treated cells received 0.001% DMSO in the medium as a vehicle. Glutathione (GSH; cat # G4251; Sigma-Aldrich) was used at a final concentration of 5 mM. Pegylated catalase (pegCAT; cat # C4963; Sigma-Aldrich) was used at a concentration of 500 U/ml. BA receptor antagonists (NF449 [95 (link)], CINPA1 [96 (link)], DY268 [97 (link)], and GSK2033 [98 (link)]) were acquired from Tocris Bioscience (Bristol, UK), and ketoconazole [99 (link)] was purchased from Sigma-Aldrich. NF449 (Gsα-selective antagonist; 5 µM) was used to inhibit TGR5 signaling. Nuclear receptor activation was inhibited using 5 µM CINPA1 (CAR antagonist), DY268 (FXR antagonist), and GSK2033 (LXR antagonist). PXR downstream signaling was inhibited using ketoconazole (5 μM). Chemotherapy drugs (irinotecan, paclitaxel, gemcitabine, 5-fluorouracil and oxaliplatin) were purchased from Sigma-Aldrich. Silencer Select siRNAs targeting TGR5 (GPBAR1-siRNA ID: s195791), VDR (NR1I1- siRNA ID: s14777), and FXR (NR1H4-siRNA ID: s19371), and the negative control siRNA #1 (cat # 4390843) were purchased from Thermo Fisher Scientific (Waltham, MA, USA) and used at a final concentration of 30 nM.
10
Cytotoxicity Assay for Anti-Cancer Agents
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Entinostat and KU-60019 were purchased from Selleck Chemicals, Munich, Germany; hydroxyurea, irinotecan, and propidium iodide (PI) were from Sigma-Aldrich Chemie GmbH, Munich, Germany; annexin V-FITC-conjugated was from Miltenyi Biotec, Bergisch Gladbach, Germany. The synthesis of COH29 was recently described in [44 (link)].
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