Female athymic nude mice aged 6–7 weeks were pre-implanted subcutaneously with pellets containing 1.7 mg β-estradiol (60-day release, Innovative Research of America, Sarasota, FL, USA). Mouse mammary fat pads were then injected with 5 × 106 MCF-7 cells stably expressing hPar1 wt, hPar1 wt and Etk/Bmx, hPar1-7A and hPar1-7A and Etk/Bmx constructs, or pcDNA3 control plasmid. Mice were monitored for tumour size by external calibre measurements (length and width) on days 10, 22, 25 and for up to 45 days if tumour burden allowed. Tumour volume (V) was calculated by V=L × W2 × 0.5, where L is length and W is width. At the end of the experiment, mice were killed and tumours were removed, weighed and fixed in formalin for histology. All animal experiments were approved by the animal committee of the Hebrew University (MD-09-11803-2).
β estradiol
Manufactured by Innovative Research
Sourced in United States
β-estradiol is a naturally occurring steroid hormone that plays a key role in the regulation of the female reproductive system. It is the primary estrogen produced by the ovaries and is essential for the development and maintenance of female secondary sexual characteristics. As a research tool, β-estradiol can be used to study the physiological and biochemical processes related to estrogen signaling and its effects on various biological systems.
Automatically generated - may contain errors
5 protocols using β estradiol
1
Modeling Breast Cancer Metastasis in Mice
2
Xenograft Tumor Model with Estrogen Implants
3
Hormone-Driven Breast Cancer Xenograft
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Female athymic nude mice were obtained from Charles River and implanted with subcutaneous, slow-release estrogen (90-day, 0.72 mg β-estradiol; Innovative Research of America). Three days after receiving the estrogen pellets, mice were injected in the fourth mammary fat pad with 2 × 107 BT-474 cells (a model of hormone receptor+/HER-2+ breast cancer), suspended in 50% Matrigel/50% RPMI1640). When tumors exceeded 95 mm3, they were eligible for randomization (8 mice/group). SDX-7320 was prepared fresh for each dose (administered s.c./Q4D × 9) in 5% mannitol/water. At necropsy, tumors were excised, and a portion snap-frozen and stored at −80°C until shipped for RNA sequencing. Methods for RNA isolation, sequencing, and analysis are described in the Supplementary Data.
4
Evaluating Anti-Tumor Effects of 5-Aza-dC
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Twelve female BALB/c nude mice, 5–6 weeks old, were obtained from the Institute of Zoology, Chinese Academy of Sciences (Beijing, China)., and were randomly divided into four groups: control (untreated, n = 3), 0.5 µM (n = 3), 5 µM (n = 3) and 10 µM (n = 3). Before injection, MCF-7 cells were maintained in the regular medium containing 5-aza-dC with the desired concentration for 72 h. On day 4, 100 µl of single cell suspensions (2.0×107 cells/ml) from untreated and treated groups were subcutaneously inoculated into lower back of nude mice. One week before inoculation, a 60-d release estrogen pellet (0.72 mg β-estradiol, Innovative Research of America) was implanted subcutaneously in each mouse. Tumor growth was evaluated by measuring the maximum diameter (A) and the minimum diameter (B) of tumor mass with a caliper at day 0, day 6, day 12, day 18, day 24 and day 30, respectively. The mean tumor volumes were calculated according to the formula V = A×B2/2. At the end of the study, mice were sacrificed by cervical dislocation and tumor masses were removed and weighed.
5
Xenograft Tumor Model with Estrogen Implants
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Estrogen pellets (14 d extended release, 0.17 mg β estradiol, Innovative Research of America) were implanted subcutaneouly into 3–4 week old female Balb/C athymic nude mice (Jackson Laboratories). Right and left inguinal mammary fatpads were injected with 106 MCF7 or MDA-MB-361 in 100 μl growth factor reduced Matrigel. Tumors were measured with calipers twice weekly. Tumor-bearing mice were randomized into treatment arms to receive fulvestrant (Novartis) delivered once weekly by intramuscular injection. All animal experimentation was performed in AAALAC approved facilities at Vanderbilt University Medical Center. All animal use protocols were reviewed and approved prior to experimentation by the Institutional Animal Care and Use Committee at Vanderbilt University, Nashville, TN.
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