Exemestane, letrozole and anastrozole were purchased from Tocris Bioscience (Bristol, UK). The activating peptide (PAR2-AP, SLIGRL-NH2) and its reverse peptide (PAR2-RP, LRGILS-NH2) of the murine PAR2 receptor were synthesized from G. Cirino (University of Naples, Naples, Italy) and dissolved in distilled water. If not otherwise indicated, all other reagents were from Sigma-Aldrich (Milan, Italy). HC-030031 was synthesized as previously described45 (link).
Letrozole
Manufactured by Bio-Techne
Sourced in United Kingdom
Letrozole is a laboratory reagent used in the field of biochemistry and cell biology research. It is a non-steroidal aromatase inhibitor that blocks the activity of the aromatase enzyme, which is responsible for the conversion of androgens to estrogens. Letrozole is commonly used in scientific research to study the role of estrogen in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anastrozole, by Bio-Techne (2 mentions)
Exemestane, by Bio-Techne (1 mentions)
Finasteride, by Merck Group (1 mentions)
17β oestradiol, by Merck Group (1 mentions)
Testosterone, by Merck Group (1 mentions)
Annexin 5, by Thermo Fisher Scientific (1 mentions)
Icatibant, by Merck Group (1 mentions)
Des arg9 leu8 bradykinin dalbk, by Merck Group (1 mentions)
Paclitaxel, by Merck Group (1 mentions)
7 aminoactinomycin d 7 aad, by BD (1 mentions)
Flutamide, by Bio-Techne (1 mentions)
Tibolone, by Merck Group (1 mentions)
7 protocols using letrozole
1
Activating murine PAR2 receptor
2
Steroid Hormone Effects on Neural Cells
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After 1 DIV the culture medium was replaced for 2 h by fresh medium devoid of B27 and GlutaMAX I supplement for Western blotting and quantitative real-time polymerase chain reaction analysis. We treated the cells for 2 h with the following test compounds, alone or in combination in the same medium: testosterone (10−7 M; Sigma-Aldrich); 17β-oestradiol (10−8 M; Sigma-Aldrich), the aromatase inhibitor letrozole (10−8 M; Tocris BioScience, Bristol, UK) or the 5α-reductase inhibitor finasteride (10−7 M; Sigma-Aldrich). The doses of steroids and inhibitors are based in previous studies13 (link)24 (link). The test compounds were dissolved in dimethyl sulfoxide and then to the final concentration in Neurobasal medium. For immunocytochemistry assays, the treatments were performed over a period of 24 h, and cultures were not deprived of B27 and GlutaMAX I supplement.
3
Modeling Bone Tumors and Lung Metastases
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To establish bone tumors, 105 SSM2 or SSM3 cells were directly injected into the tibia of WT male or female mice. When indicated, female mice were either SHAM-operated or ovariectomized (OVX) under general anesthesia. 105 SSM2 or SSM3 cells were intratibially injected either one week post-surgery or two weeks pre-surgery. Animals were fed an estrogen-free diet throughout the duration of the experiments (Harlan Teklad, Madison, WI, USA). In some experiments, animals received the aromatase inhibitor Letrozole (TOCRIS Bioscience) at 10 ug/day in addition to OVX surgery, for 4 weeks.
To establish lung metastases, 5 × 105 SSM2 cells were injected into the tail vain of WT male mice. Another group of WT male mice received 105 SSM2 cells into the right tibias and were used as control for tumor growth. Animals in both groups were euthanized 30 days post tumor inoculation.
To establish lung metastases, 5 × 105 SSM2 cells were injected into the tail vain of WT male mice. Another group of WT male mice received 105 SSM2 cells into the right tibias and were used as control for tumor growth. Animals in both groups were euthanized 30 days post tumor inoculation.
4
Pharmacological Modulation of Bradykinin Receptors
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Anastrozole and letrozole were purchased from Tocris Bioscience (Bristol, UK) and were diluted in 0.5% carboxymethylcellulose and 99.5% of NaCl (0.9%). Paclitaxel (6 mg/mL of Paclitaxel in Cremophor EL and dehydrated ethanol) was purchased from Glenmark (Buenos Aires, ARG) and was dissolved in NaCl (0.9%). Icatibant (kinin B2 receptor peptide antagonist), des-Arg9-[Leu8]-bradykinin (DALBk; kinin B1 receptor peptide antagonist) were purchased from Sigma-Aldrich Chemical Company (St. Louis MO, USA) and were also prepared in NaCl (0.9%). The enzyme immunoassay kit for bradykinin was obtained from Phoenix Pharmaceuticals, Inc. (California, USA). Specific anti-B1 (bs-8675R–lot 9C20V14) or anti-B2 (bs-2422R–lot AG08307921) antibodies were acquired from Bioss Antibodies (Massachusetts, USA) and secondary antibody (sc-2357–lot L1218) was obtained from Santa Cruz Biotechnology (California, USA). Paclitaxel from semisynthetic ≥ 97% (in vitro assays) was acquired from Sigma-Aldrich Chemical Company (St. Louis MO, USA). Annexin-V and 3- (4, 5-dimethyl-2-thiazolyl) -2, 5-diphenyl-2H-tetrazolium bromide] salt (MTT; 5 mg/mL solution) were obtained from Life Technologies (São Paulo, BR). The 7-Amino-Actinomycin D (7AAD) was acquired from BD Biosciences (California, USA). The doses of the drugs used in this study were based on previous studies13 (link),15 (link),23 (link),40 (link),72 (link).
5
Astrocyte Phagocytosis Modulation by Steroid Receptors
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Astrocytes were incubated for 6 h in DMEM with 10% FBS and then 15 h with the following test compounds, either alone or in combination in DMEM 0.1% FBS: tibolone (100 nM; Sigma-Aldrich); ERα antagonist 1,3-Bis(4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidyletoxy) phenol]-1H-pyrazole (MPP, 100 nM; Sigma-Aldrich); the ERβ antagonist 4-[2-phenyl-5,7-bis (trifluoromethyl) pyrazolo [1,5-a] pyrimidin-3-yl] phenol (PHTPP, 100 nM; Tocris, Bristol); the G-protein-coupled ER (GPER) antagonist (3aS,4R,9bR)-4-(6-bromo-1,3-benzodioxol-5-yl)-3,4,5,9B-tetrahydro-3H-cyclopenta[c]quinolone (G15, 100 nM; Tocris); the androgen receptor antagonist flutamide (100 nM; Tocris); or the aromatase inhibitor letrozole (100 nM; Tocris). Twenty-four hours before phagocytosis assay astrocytes were incubated with DMEM serum-free medium with the previous compounds plus lipopolysaccharide (LPS) isotype O26:B6 (1 μg/ml; Sigma-Aldrich, Tres Cantos, Madrid). The concentrations used for the test compounds were chosen on the basis of previous studies [11 (link), 42 (link)]. After the incubation with the test compounds, phagocytosis activity was assessed as indicated below.
6
Letrozole Dosage for Delayed Implantation
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In order to explore the optimal dose of delayed implantation, letrozole (Tocris) was dissolved in DMSO in a dose of 0.1, 0.5, 0.8, 1, 2, and 5 mg/kg body weight, respectively. Mice were injected intraperitoneally with different dose of letrozole at 1200 h on day 3 of pregnancy, and control mice were injected with 50 μl DMSO alone.
Implantation sites were checked on day 5 by intravenous blue dye injection. In order to exclude the situation of failed fertilization, uteri from these mice with no blue dye reaction were collected and flushed with 0.9% saline water. Only the mice with embryos were counted as delayed implantation.
In the 0-day delayed model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 or 25 ng E2 was injected on the morning of day 4 to activate embryo implantation.
In the 1-day delay model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 was injected subcutaneously on the morning of day 5 to activate embryo implantation.
Implantation sites were checked on day 5 by intravenous blue dye injection. In order to exclude the situation of failed fertilization, uteri from these mice with no blue dye reaction were collected and flushed with 0.9% saline water. Only the mice with embryos were counted as delayed implantation.
In the 0-day delayed model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 or 25 ng E2 was injected on the morning of day 4 to activate embryo implantation.
In the 1-day delay model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 was injected subcutaneously on the morning of day 5 to activate embryo implantation.
7
Letrozole Dosage for Delayed Implantation
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In order to explore the optimal dose of delayed implantation, letrozole (Tocris) was dissolved in DMSO in a dose of 0.1, 0.5, 0.8, 1, 2, and 5 mg/kg body weight, respectively. Mice were injected intraperitoneally with different dose of letrozole at 1200 h on day 3 of pregnancy, and control mice were injected with 50 μl DMSO alone.
Implantation sites were checked on day 5 by intravenous blue dye injection. In order to exclude the situation of failed fertilization, uteri from these mice with no blue dye reaction were collected and flushed with 0.9% saline water. Only the mice with embryos were counted as delayed implantation.
In the 0-day delayed model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 or 25 ng E2 was injected on the morning of day 4 to activate embryo implantation.
In the 1-day delay model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 was injected subcutaneously on the morning of day 5 to activate embryo implantation.
Implantation sites were checked on day 5 by intravenous blue dye injection. In order to exclude the situation of failed fertilization, uteri from these mice with no blue dye reaction were collected and flushed with 0.9% saline water. Only the mice with embryos were counted as delayed implantation.
In the 0-day delayed model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 or 25 ng E2 was injected on the morning of day 4 to activate embryo implantation.
In the 1-day delay model, delayed implantation was induced by injection with letrozole at a dose of 5 mg/kg body weight on the day 3 of pregnancy, and 3 ng E2 was injected subcutaneously on the morning of day 5 to activate embryo implantation.
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