Two vitamin D compounds PRI-2191 and PRI-2205 obtained at the Chemistry Department of the Pharmaceutical Research Institute, Warsaw, Poland were used. Calcitriol was used as a reference standard for both analogs. Samples of the compounds were stored in amber ampoules under argon at −20 °C. Directly before use, the compounds were dissolved in absolute ethanol to achieve the concentration of 100 µM and subsequently diluted in a culture medium to reach the required concentrations (10 nM for T47D or 100 nM for other cancer cell lines). For animal experiments, the compounds were dissolved in 99.8% (v/v) ethanol, diluted in 80% (w/v) propylene glycol to reach the required concentrations, and administered s.c. to mice at the volume of 5 µL/g of animal b.w. Anastrozole (Sigma-Aldrich, Steinheim, Germany), and was dissolved in 40% (v/v) ethanol (10 mg/mL) and subsequently diluted in a culture medium to reach the required concentration of 100 μg/mL. For animal experiments, Anastrozole was dissolved in 40% (v/v) ethanol and then diluted in 0.9% (w/v) saline to reach the concentration of 0.2 mg/animal/0.1 mL.
Anastrozole
Manufactured by Merck Group
Sourced in United States, Germany
Anastrozole is a type of laboratory equipment used in scientific research and experiments. It is a selective aromatase inhibitor, which means it helps to inhibit the activity of the aromatase enzyme. This enzyme is responsible for the conversion of androgens to estrogens. Anastrozole is commonly used in various research applications, but its core function is to help control and regulate the levels of estrogen in experimental settings.
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Lab products found in correlation
4 androstene 3 17 dione, by Merck Group (3 mentions)
Formic acid, by Merck Group (2 mentions)
Letrozole, by Merck Group (2 mentions)
Estradiol, by Merck Group (2 mentions)
Progesterone, by Merck Group (2 mentions)
Ru486, by Merck Group (2 mentions)
Dihydrotestosterone, by Steraloids (2 mentions)
17β estradiol, by Merck Group (2 mentions)
Testosterone, by Merck Group (2 mentions)
Doxorubicin, by Merck Group (2 mentions)
Cisplatin, by Merck Group (2 mentions)
Exemestane, by Merck Group (2 mentions)
Dutasteride, by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Scintillation cocktail, by Merck Group (1 mentions)
Testosterone d3, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Acebutolol, by Merck Group (1 mentions)
19 protocols using anastrozole
1
Vitamin D Analogs and Antiestrogen Evaluation
2
Compound Dilution Protocol for Drug Treatments
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Dutasteride and anastrozole were obtained from Sigma-Aldrich (St. Louis, MO, USA). Finally, ASP9521 was obtained from Adooq bioscience (Irvine, CA, USA). All drugs were diluted in dimetil-sulfoxide (DMSO) (Sigma Aldrich, St. Louis, MO, USA) to start from an initial concentration of 20 mM, 34 mM and 30 mM for Dutasteride, anastrozole and ASP9521, respectively. Then, drugs were diluted again to achieve a work concentration of 10 mM and 1 mM for all treatments.
3
Steroid Hormone Extraction and Analysis
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Dexamethasone (internal standard), 4-Androstene-3,17-dione-2,3,4-13C3, Estrone-2,3,4-13C3, 17β-Estradiol-2,3,4-13C3, Testosterone-2,3,4-13C3, Anastrozole, Letrozole, KZ, protease from Bacillus licheniformis (subtilisin), formic acid, acetone, methanol, acetonitrile, hexane, methylene chloride, charcoal activated, Tris, HCl, KH2PO4, K2HPO4, KCl, NADPH, glycerol, BSA, scintillation cocktail, and Supel™-Select SPE HLB (six mL, 200 mg) columns were provided by Sigma-Aldrich (St. Louis, MO, USA). 1β-3H Androstene-3,17-dione (30 Ci/mmol) was obtained from Perkin-Elmer Life Science (Boston, MA, USA).
4
Breath Sampling Protocol for Doping Substances
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Breath sampling units were obtained from SensAbues (Huddinge, Sweden). Methanol, acetonitrile, formic acid, and ammonium acetate were of analytical grade and purchased from Merck (Darmstadt, Germany). The model substances were provided by different suppliers: bisoprolol dehydrochloromethyltestosterone (DHCMT), meldonium, metoprolol, methylhexaneamine, and pseudoephedrine were from LGC Standards GmbH (Wesel, Germany). Acebutolol, (S)‐2‐aminooctane, anastrozole, letrozole, methylphenidate, stanozolol, D3‐testosterone, and D7‐propranolol were from Sigma (Schnelldorf, Germany). D3‐Meldonium was from Toronto Research Chemicals (Toronto, Canada), and YK‐11 was synthesized in‐house as described elsewhere.
5
Hormonal Manipulation in Mouse Models
6
Aromatase Inhibitor and Growth Hormone Effects
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Anastrozole, a selective non-steroidal aromatase inhibitor (AI), 17-β estradiol, testosterone, flutamide and Oil Red O stain were purchased from Sigma-Aldrich Co., Bangalore, India. Recombinant bovine growth hormone (rbGH) was a kind gift from Monsanto Company St. Louis, MO. Oligonucleotide and Oligo dT primers were synthesized by Sigma-Genosys, Bangalore, India. DyNAzyme™ II DNA polymerase was purchased from Finnzymes (Espoo, Finland) and dNTPs were procured from Eppendorf, (Hamburg, Germany). Power SYBR® Green PCR master mix was obtained from Applied Biosystems, Foster City, CA. The details of antibodies employed are provided in Additional file 1 : Table S1. The secondary anti-rabbit and the ABC colour development kit was procured from Bangalore Genei, India. Mouse/rat Insulin-like Growth Factor-1 (m/r IGF-1) ELISA kit was procured from BioVendor, Mediagnost, Germany. All other reagents were purchased from Sigma-Aldrich Co., Bangalore, India or sourced from local distributors.
7
Hormonal Manipulation in Mouse Models
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Specific mouse alleles used in this study are referenced in full Methods. Mice were housed in AAALAC-accredited, specific-pathogen-free animal care facilities at the University of Michigan (UM), Baylor College of Medicine (BCM), or UT Southwestern Medical Center (UTSW). All procedures were approved by the UM, BCM, and UTSW Institutional Animal Care and Use Committees. For hormonal treatment, mice were injected subcutaneously with 100 μl of corn oil containing 2 μg estradiol (Sigma, St. Louis, MO), 1 mg progesterone (Sigma)5 (link), or 100 μg of dihydrotestosterone (Steraloids, Newport, RI). 50 μg of anastrozole (Sigma) dissolved in PBS was given intraperitoneally. RU486, PPT, and DPN (all from Sigma) dissolved in corn oil were administered subcutaneously at 5mgkg−1. pIpC (Amersham, Piscataway, NJ) was resuspended in PBS at 50 μg/ml, and 0.5 μg/gram of body mass were injected intraperitoneally every other day for 6 days. Females were mated with male mice one week after the last injection.
8
Breast Cancer and Adipocyte Interactions
9
Breast Cancer Treatment Compound Evaluation
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17β-estradiol (E2), 4-OHT, anastrozole, insulin, hydrocortisone, cholera toxin, and DMSO were purchased from Sigma (St. Louis, MO). EGF was purchased from Peprotech (Rocky Hill, NJ), horse serum was from Invitrogen (Carlsbad, CA). TAM pellets (5 mg/pellet, 60-day release) and E2 pellets (0.36 mg/pellet, 60-day release) were purchased from Innovative Research of America (Sarasota, FL, USA). Matrigel™ Matrix Growth Factor Reduced was purchased from BD Biosciences (Bedford, MA, USA). Anti-Raf-B, anti-p21, anti-Bcl-2, anti-Fibronectin and anti-β-actin antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). BreastDefend® (BD) was supplied by EcoNugenics, Inc. (Santa Rosa, CA, USA) and dissolved in DMSO at a concentration of 25 mg/ml then stored at −20 °C. The chemical composition of BD was previously published [26 (link)]. All other chemicals and reagents were of analytical grade.
10
Glioblastoma Xenograft Treated with Anastrozole
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We formed two groups of mice; both groups received a C6 cells’ xenograft; the first group was not treated (GBM group, n = 5), and the other one was treated with Anastrozole (GBM+Anastrozole, n = 5). We anesthetized mice with sevoflurane (3%). We made an incision in the brain midline of the scalp and a small hole in the skull following the stereotaxic coordinates (X = 1.34 mm, Y = 1.5 mm, and Z = 3.5 mm). We administered 1 × 106 cells in 2 μL of DMEM-F12 using a Hamilton syringe in mice’s right striatum (See Supplementary Materials ). Anastrozole (Sigma Aldrich A2736, MO, USA) was dissolved in DMSO 0.1 mM to obtain a final concentration of 500 μg/mL (stock solution) and stored at −20 °C. The drug (0.1 mg/kg) was administered through the tail vein with an insulin syringe (0.5 mL daily) for seven days.
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