BO-1055 was synthesized at Dr. Tsann Long Su's laboratory at the Institute of Biomedical Sciences (IBMS), Academia Sinica, Taipei, Taiwan. PU-H71 was synthesized at Dr. Gabriela Chiosis' laboratory at Memorial Sloan-Kettering Cancer Center (MSKCC), New York, NY, USA. Topotecan was purchased from Sigma-Aldrich (Saint Louis, MO), etoposide, melphalan and 4-hydroperoxy cyclophosphamide (4-HC) were purchased from Santa Cruz Biotechnology (Dallas, TX). SN38, Bendamustine hydrochloride, vincristine sulfate and cisplatin were purchased from Tocris (Bio-Techne, Minneapolis, MN). Doxorubicin and cyclophosphamide were purchased from the pharmacy at MSKCC.
Sn 38
Manufactured by Bio-Techne
Sourced in United Kingdom
SN-38 is a chemical compound that serves as an active metabolite of the anti-cancer drug irinotecan. It functions as a topoisomerase I inhibitor, which is a crucial mechanism for disrupting cancer cell division and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Irinotecan, by Merck Group (3 mentions)
Cisplatin, by Bio-Techne (2 mentions)
Rabeprazole, by Santa Cruz Biotechnology (2 mentions)
Hct116, by American Type Culture Collection (2 mentions)
Ht 29, by American Type Culture Collection (2 mentions)
Topotecan, by Merck Group (1 mentions)
Etoposide, by Santa Cruz Biotechnology (1 mentions)
Melphalan, by Santa Cruz Biotechnology (1 mentions)
4 hydroperoxy cyclophosphamide, by Santa Cruz Biotechnology (1 mentions)
Vincristine sulfate, by Bio-Techne (1 mentions)
Cell proliferation kit 2, by Roche (1 mentions)
Oxaliplatin, by Bio-Techne (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Lps o55 b5, by Merck Group (1 mentions)
Doxorubicin, by Bio-Techne (1 mentions)
Torin, by Cell Signaling Technology (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Ls174t, by American Type Culture Collection (1 mentions)
Salinomycin, by Merck Group (1 mentions)
Abt 737, by Selleck Chemicals (1 mentions)
13 protocols using sn 38
1
Synthesis and Procurement of Cancer Drugs
2
Cellular Viability and SN38 IC50 Determination
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Cellular viability and IC50 for SN38 were determined using the Cell Proliferation Kit II (XTT) according to the manufacturer's instructions (Roche Applied Science, Indianapolis, IN). The cells were seeded at a density of 5000 cells/cm2 in 96 well plates and allowed to attach for 24 hr. Freshly prepared SN38 (Tocris Bioscience, UK) was then added with a final concentration ranging from 0 to 1000 nM. After 48 hr of treatment cellular viability was determined using the XTT assay. The assay is based on the cleavage of the yellow tetrazolium salt XTT (2, 3-Bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carbox-anilide) into a soluble orange formazan dye. This reaction is attributed mainly to the succinate-tetrazolium reductase system in the mitochondria of metabolically active cells. The absorbance measured at 450 nm is proportional to the number of viable cells. Experiments were repeated two times independently for cytokine and chemokine analysis.
3
Viability Assay for Chemotherapeutic Evaluation
4
Colon Cancer Cell Lines Treatment
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The human colon cancer cell lines HCT116, HT29, DLD1, and LoVo were obtained from ATCC. HCT116 and DLD1 cells were grown and maintained in RPMI containing 10% FBS and pen-strep. HT29 cells were grown and maintained in McCoy’s medium containing 10% FBS and pen-strep. LoVo cells were grown and maintained in Ham’s F12-K containing 10% FBS and pen-strep. All cells were grown at 37 °C and 5% CO2 in a humidified cell culture incubator. Topo I inhibitor treatment was performed using various concentrations of either irinotecan (Sigma-Aldrich) or SN-38 (Tocris). SN-38 is the irinotecan active metabolite. Cells were also treated with various concentrations of Rabeprazole (Santa Cruz Biotechnology).
5
Reconstitution of TLR4 Agonists and Antagonists
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TLR4 antagonists TAK-242 (Sapphire Bioscience, Australia) and IAXO-102 (Innaxon, UK) and TLR4 agonists and inflammatory mediators, LPS O55:B5 (Sigma-Aldrich, USA), SN-38 (Tocris Bioscience, United Kingdom) and TNF-α (Research and Diagnostic Systems, United States), were reconstituted according to manufacturer’s instructions for in vitro and ex vivo experiments: TAK-242: DMSO; IAXO-102: DMSO and ethanol; LPS: sterile MilliQ water; SN-38: DMSO; TNF-α: sterile PBS and 0.1% bovine serum albumin.
6
Senescence Induction and Escape Dynamics
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MCF-7 and LS174T cell lines were obtained from the American Type Culture Collection. Cell lines were authenticated by STR profiling and regularly tested to exclude mycoplasma contamination. To induce senescence, MCF-7 and LS174T cells were treated in RPMI medium (Dutscher; cat. no. L0500-500) containing 3% fetal bovine serum (FBS) (Eurobio; cat. no. CVFSVF00 01) respectively with doxorubicin (Tocris Bioscience; cat. no. 2252) (25 ng/ml) and sn38 (Tocris Bioscience; cat. no. 2684) (5 ng/ml) for 96 h. To promote senescence escape, cells were washed with PBS and stimulated with fresh medium containing 10% FBS for the indicated time. AKT inhibitor [iAKT1/2:1,3-Dihydro-1-(1-[(4-[6-phenyl-1H-imidazo(4,5-g)quinoxalin-7-yl]phenyl)methyl]-4-piperidinyl)-2H-benzimidazol-2-one trifluoroacetate; Sigma-Aldrich; Merck KGaA; cat. no. A6730] was used at a concentration of 100 µM and Torin (Cell Signaling Technology, Inc.; cat. no. 14379) at 10 nM.
7
Synthesis and Preparation of Plocabulin
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8
Persistent Colorectal Cancer Cell Lines
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Colorectal cell lines (American Type Culture Collection) were maintained in antibiotic-free RPMI 1640 medium (Lonza), supplemented with 10% fetal bovine serum and maintained at 37 °C in 5% carbon dioxide. Cells were routinely tested for the absence of mycoplasma contamination. Unless indicated, note that cell treatments were performed in 3% FBS. For persistent cell generation, cells were treated for the indicated times with sn38 (5ng/ml; Tocris Bioscience 2684) in 3% FBS, washed with PBS and then restimulated with fresh 10% FBS. Proliferation generally resumed after 3-5 days. Other treatments were used : Salinomycin (1 or 5μM; Sigma Aldrich S6201) and ABT737 (1μM; Selleckchem S1002).
9
Formulation and Evaluation of SN-38 Lipid Nanoparticles
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L-α-distearoyl-phosphatidylcholine (DSPC), egg yolk phosphatidylcholine (EPC), soy bean lipid extract (SLE), L-α-dioleoyl-phosphatidylserine (DOPS), and cholesterol (CHOL) were purchased from Avanti Polar Lipids (Birmingham, AL, USA). SN-38 was from Tocris Bioscience (Bristol, UK). All the organic solvents (Panreac, Montcada y Reixac, Barcelona, Spain) were distilled before use. Milli-Q water (resistivity 18 MΩ⋅cm; Millipore, Bedford, MA, USA) was used. All other chemicals and solvents were of analytical grade. Dulbecco’s Modified Eagle’s Medium (DMEM) with 4.5 g glucose/L, fetal calf serum, L-glutamine, and penicillin–streptomycin solutions were provided by Biological Industries (Crowell, CT, USA). Sterile Dulbecco’s phosphate buffered saline (PBS), DMSO, 3-[4,5-dimethylthiazol-2-yl] 2,5-diphenyltetrazolium bromide (MTT), Trypan blue, toluidine blue, and Hoechst-33258 (H-33258) were from Sigma-Aldrich Chemical Co. (St Louis, MO, USA). The CytoScan-Fluoro Assay Kit was provided by G-Biosciences (St Louis, MO, USA). Sterilized plastic material was purchased from Corning Incorporated (Corning, NY, USA).
10
Mitoxantrone Transport and Resistance Assay
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