Salinomycin

Manufactured by Cayman Chemical

Sourced in United States

Salinomycin is a polyether ionophore antibiotic commonly used as a laboratory reagent. It functions as a cation exchanger and can transport monovalent cations across cell membranes.

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5 protocols using salinomycin

Dual-Color Bioluminescence Imaging for Growth and Cytotoxicity

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For growth and cytotoxicity curves, we performed dual-dolor bioluminescence imaging of CBRed and CBG using an IVIS Lumina Series III (Perkin Elmer, Waltham, MA, USA) as previously described (34 (link)). We exported photon flux data from regions of interest defined in Living Image 4.3.1 (Perkin Elmer) and quantified changes in growth or viability with custom MATLAB code.
For cytotoxicity curves, we treated cells with compounds diluted in LG/LF media for three days. We used ferric ammonium citrate (FAC) (#F5879, Sigma Aldrich), fulvestrant (#S1191, Selleckchem), tamoxifen, deferoxamine (DFO), deferasirox (DFX), RSL-3, salinomycin, and erastin (#13258, #14595, #16753, #17754, #19288, and #13579, Cayman Chemical). We manufactured salinomycin analogs AM5 and AM23 as previously described (35 ).

Buschhaus J.M., Rajendran S., Humphries B.A., Cutter A.C., Muñiz A.J., Ciavattone N.G., Buschhaus A.M., Cañeque T., Nwosu Z.C., Sahoo D., Bevoor A.S., Shah Y.M., Lyssiotis C.A., Ghosh P., Wicha M.S., Rodriguez R, & Luker G.D. (2022). Effects of iron modulation on mesenchymal stem cell-induced drug resistance in estrogen-receptor-positive breast cancer. Oncogene, 41(29), 3705-3718.

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Screening Anti-Cancer Stem Cell Agents

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We cultured HCT116, HT29, and DLD-1 cells under standard culture conditions in RPMI-1640 media (Welgene) containing 10% FBS (Atlas Biologicals) and 1 × penicillin-streptomycin (Corning). For the drug treatments, we dissolved JIB-04 (Cayman) and salinomycin (Cayman) in DMSO and diluted the solutions to the appropriate concentrations for the experiments. The following chemicals were used for the primary screening of anti-CSC agents: IOX1 (Cayman), SAHA (Cayman), trichostatin (TSA) (Cayman), EPZ5676 (APEXBIO), eosin Y disodium trihydrate(AMI-5) (Santa Cruz), 2-PCPA(Cayman), sirtinol (Cayman), pargyline (Sigma-Aldrich), paclitaxel (Cayman) 5-FU (Cayman), etoposide (Cayman), nocodazole (Sigma-Aldrich), calmidazolium chloride (Tocris Bioscience), beta-lapachone (Cayman).

Kim M.S., Cho H.I., Yoon H.J., Ahn Y.H., Park E.J., Jin Y.H, & Jang Y.K. (2018). JIB-04, A Small Molecule Histone Demethylase Inhibitor, Selectively Targets Colorectal Cancer Stem Cells by Inhibiting the Wnt/β-Catenin Signaling Pathway. Scientific Reports, 8, 6611.

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Antitumor Effects of Drug Treatments on Xenograft Tumors

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ATG5‐WT or ATG5‐KO cells were mixed with NSPC medium and Matrigel matrix (Corning, Cambridge, MA, USA) (1:1 ratio) at a concentration of 1 × 10⁴ cells/μL. Cells (1 × 10⁶ cells/100 μL) were subcutaneously transplanted into each of the 2 flanks of anesthetized female Balb/c nu/nu mice. Transplanted mice were randomly distributed into groups (4 mice/group) for drug treatments: control (DMSO, every day); TMZ (Sigma‐Aldrich) (0.1 mg/kg/d, every day); nigericin (Sigma‐Aldrich) (1 mg/kg/d, every day); or salinomycin (Cayman Chemical, Michigan, USA) (5 mg/kg/d, every 2 days). Drugs dissolved in DMSO were mixed with corn oil and intraperitoneally (ip) injected 1 day after transplantation. CQ dissolved in 20% DMSO/PBS(−) were injected (ip 25 mg/kg/d, every day). All drugs were injected into mice for 2 weeks starting on day 1 after transplantation. The widths (W) and lengths (L) of tumors developing in recipients were measured using calipers. Tumor volumes were calculated using the formula: volume (V) = (W² × L)/2. For intracranial glioma cell injections, cells (1 × 10⁵/mouse) were transplanted into anesthetized 4‐week‐old female Balb/c nu/nu mice as described previously.11 All animal experiments were approved by the Committee on Animal Experimentation of Kanazawa University and performed following the university's guidelines for the care and use of laboratory animals.

Vu H.T., Kobayashi M., Hegazy A.M., Tadokoro Y., Ueno M., Kasahara A., Takase Y., Nomura N., Peng H., Ito C., Ino Y., Todo T., Nakada M, & Hirao A. (2018). Autophagy inhibition synergizes with calcium mobilization to achieve efficient therapy of malignant gliomas. Cancer Science, 109(8), 2497-2508.

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Cell Signaling Pathway Modulators

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Nonactin, valinomycin, carbonyl cyanide m‐chlorophenyl hydrazone (CCCP), and 2‐deoxyglucose (2‐DG), Z‐VAD‐FMK were purchased from Sigma‐Aldrich (St. Louis, MO). Salinomycin and monensin were purchased from Cayman Chemical (Ann Arbor, MI). Wnt‐3a ligand was purchased from Wako Pure Chemical Industries (Osaka, Japan).

Shikata Y., Kiga M., Futamura Y., Aono H., Inoue H., Kawada M., Osada H, & Imoto M. (2017). Mitochondrial uncoupler exerts a synthetic lethal effect against β‐catenin mutant tumor cells. Cancer Science, 108(4), 772-784.

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Synthesis and Characterization of Ophiobolin Analogues

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Curcumin, genestein, doxorubicin, and paclitaxel were obtained from Selleckchem (Houston, TX, USA), salinomycin from Cayman Chemicals (Ann Arbor, MI USA), and disulfiram from Tocris Bioscience, (Bristol, UK). OpA was produced by fermentation of the fungus D. gigantea. It was extracted from the fungal culture filtrates, purified, crystallized and identified by ¹H NMR and ESI MS spectra as previously reported^{70 (link)}. The purity of OpA was > 98% as ascertained by ¹H NMR and HPLC analyses.
3-Deoxy OpA was synthesized from ophiobolin I^{71 (link),72 (link)} which was also obtained through fermentation as previously reported^{70 (link)}. A two-step synthetic sequence involving conjugate reduction of the enone which proceeded with high diastereoselectivity (> 19:1 by 600 MHz ¹H NMR) followed by a Ru(IV)-mediated oxidation of the primary alcohol to the aldehyde delivered 3-deoxy OpA. It should be noted that the methyl group at C3 is epimeric with respect to the C3-methyl group in OpA. However, the importance of the C3-hydroxy group and/or the stereochemistry of this methyl group was verified through studies described below and 3-deoxy OpA served as a negative control. Further details are provided in Supplemental Figure 5.

Reisenauer K.N., Tao Y., Das P., Song S., Svatek H., Patel S.D., Mikhail S., Ingros A., Sheesley P., Masi M., Boari A., Evidente A., Kornienko A., Romo D, & Taube J. (2021). Epithelial-mesenchymal transition sensitizes breast cancer cells to cell death via the fungus-derived sesterterpenoid ophiobolin A. Scientific Reports, 11, 10652.

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