Treatment was initiated 2 to 3 weeks (dose-response study) or at day 20 (combination study) after cancer cell injections. For the dose-response study, LAS at 1, 5, or 10 mg/kg in 100 μL of PBS containing 15% PEG 400, or vehicle, was administered 5 days/week via subcutaneous (s.c.) injection, while FUL 5 mg/mouse (Med Chem Express Cat#HY-13636) in 100 μl of mineral oil was injected s.c. once per week. For the combination study, PAL (Med Chem Express Cat#HY-50567) at 35 or 70 mg/kg in 100 μL of 50 mM sodium lactate (pH4) was administered via oral gavage 5 days/week either alone or in combination with LAS or FUL. Mice were sacrificed 70 days after treatment initiation (dose-response study) or 82 days after cancer cell injection (combination study), and mammary gland tumors were weighed. Metastases were evaluated with ex vivo imaging of excised long bones, brains, lungs, and livers and quantified by luciferase activity for the combination study.
Hy 13636
Manufactured by MedChemExpress
Sourced in United States
HY-13636 is a piece of laboratory equipment manufactured by MedChemExpress. It is designed to perform specific tasks in a research or analytical setting. The core function of this product is to aid in the execution of scientific experiments or data collection, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Hy 50567, by MedChemExpress (2 mentions)
Hy 19424, by MedChemExpress (1 mentions)
Hy 103456, by MedChemExpress (1 mentions)
Hyclone, by GE Healthcare (1 mentions)
Collagenase dispase, by Merck Group (1 mentions)
Af 100 18b, by Thermo Fisher Scientific (1 mentions)
Fibroblast growth factor (fgf), by Thermo Fisher Scientific (1 mentions)
Af 100 15, by Thermo Fisher Scientific (1 mentions)
Accutase, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Cholera toxin, by Merck Group (1 mentions)
Celltiter glo 3d reagent, by Promega (1 mentions)
4 protocols using hy 13636
1
Dose-Dependent and Combination Therapy for Breast Cancer
2
Investigating Estrogen and Heme Effects
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The Ishikawa and RL95-2 cells were treated with estrogen (10−7 M, HY-B070, MedChem Express, NJ, USA), succinate (5 mM, Sigma-Aldrich; Merck KGaA, SL, MO, USA), ERα antagonist (MPP, 10−6 M, HY-103454, MedChem Express, Monmouth Junction, NJ, USA), ERβ antagonist (PHTPP, 10−6 M, HY-103456, MedChem Express, Monmouth Junction, NJ, USA), total ER antagonist Fulvestrant (ICI182780, 10−6 M, HY-13636, MedChem Express, Monmouth Junction, NJ, USA), and heme (0–100 μM, HY-19424, MedChem Express, Monmouth Junction, NJ, USA).
Both cell lines were seeded in 12-well plates at a density of 2.5 × 105 cells/mL and cultured in DMEM/F12 medium (HyClone; GE Healthcare Life Sciences) supplemented with 10% FBS-charcoal-stripped (Gibco Cell Culture, Carlsbad, CA, USA) and 1% penicillin–streptomycin (HyClone, UT, USA). The relevant cells were cultured under 5% CO2 in a 37 °C humidified incubator (Heal Force HF-90, Shanghai, China) for 48 h.
Both cell lines were seeded in 12-well plates at a density of 2.5 × 105 cells/mL and cultured in DMEM/F12 medium (HyClone; GE Healthcare Life Sciences) supplemented with 10% FBS-charcoal-stripped (Gibco Cell Culture, Carlsbad, CA, USA) and 1% penicillin–streptomycin (HyClone, UT, USA). The relevant cells were cultured under 5% CO2 in a 37 °C humidified incubator (Heal Force HF-90, Shanghai, China) for 48 h.
3
Evaluating Combination Therapy for Tumor Growth
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Mice were randomized to vehicle, LAS, FUL, PAL, LAS plus PAL, or FUL plus PAL (6 mice/group) 2-3 weeks after cell injections. Vehicle and LAS (10 mg/kg in 100 µL of PBS containing 15% PEG400) were administered subcutaneously 5 days/week, FUL (Med Chem Express, HY-13,636; 5 mg/mouse in 100 µL of mineral oil) subcutaneously once per week, and PAL (Med Chem Express, HY-50,567; 70 mg/kg in 100 µL of 50 mM sodium lactate buffer pH 4) via oral gavage 5 times/week. Clinically, LAS and FUL are administered orally and intramuscularly. However, for this study we injected both LAS and FUL subcutaneously for convenience; while that may be considered a limitation, equivalent biological responses with oral and subcutaneous administration of LAS and other SERMs were observed in preliminary, unpublished, preclinical dosing animal experiments. This experiment was repeated with the same design and 8-9 mice in each treatment group.
4
Rat Mammary Tumor Organoid Assay with Fulvestrant
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Rat mammary tumor and autologous healthy kidney tissues were minced into 1 to 2 mm fragments then digested with 1 mg/mL collagenase/dispase (Sigma, 11097113001) for 30 minutes. Digestion was stopped by adding equal volume of 1% BSA in DMEM, then centrifuged at 1,500 rpm × 5 minutes. Pellets were further digested with Accutase (Sigma, A6964) for 30 minutes then collected by centrifugation at 1,500 rpm for 5 minutes. Pellets were then resuspended in organoid growth medium containing Y-27632, 5% matrigel, and growth factors including B27 (Thermo Fisher Scientific, 1750404), insulin (Sigma, 12643), cholera toxin (Sigma, C8052), FGF (Peprotech, AF-100-18B), EGF (Peprotech, AF-100-15), and IL6 (Peprotech, 200-06) The suspension was seeded onto 12-well plates precoated with matrigel. Culture media was replaced every 4 days. For fulvestrant experiments, organoids were plated at 5,000 to 10,000 single cells per well on top of a layer of matrigel in 96-well plates. On day 6, organoids were treated with fulvestrant (MedChem Express, HY-13636; 100 and 750 nmol/L) or DMSO vehicle control. After 72 hours of treatment, CellTiter-Glo 3D reagent (Promega, G9682) was added to each well and luminescence signal was detected after 30 minutes using a Synergy H1 microplate reader.
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