Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Estrogen Modulates Cartilage and Subchondral Bone Remodeling in an Ovariectomized Rat Model of Postmenopausal Osteoarthritis

doi: 10.12659/MSM.916254

Figure Lengend Snippet: Three-dimensional (3D) micro-computed tomography (micro-CT) images of the subchondral bone in the ovariectomized/destabilization of the medial meniscus (OVX/DMM) rat group with and without estrogen (E2) and raloxifene (RAL) treatment and the control group. ( A ) Severe bone erosion, cyst formation, and bone sclerosis are shown in the OVX/DMM group. ( B ) Near-normal subchondral bone in the OVX/DMM+E2 group. ( C ) The subchondral bone shows osteoporosis and the bone structure is destroyed in the OVX/DMM+RAL group. ( D–F ) The degree of osteoporosis in the subchondral bone was increased.

Article Snippet: Two months after the models were successfully established, rats were injected with estrogen (0.1mg/kg/day) (Sigma-Aldrich, St. Louis MO, USA) or raloxifene (3mg/kg/day) (MedChem Express, Monmouth Junction, NJ, USA) for one month.

Techniques: Micro-CT, Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Estrogen Modulates Cartilage and Subchondral Bone Remodeling in an Ovariectomized Rat Model of Postmenopausal Osteoarthritis

doi: 10.12659/MSM.916254

Figure Lengend Snippet: The bone mineral density (BMD) of the subchondral bone at 0, 2, and 4 weeks after surgery. Compared with the control group, the ovariectomized/destabilization of the medial meniscus (OVX/DMM) rat group, the OVX/DMM+estrogen (E2) group, and the raloxifene (RAL) treated group all show a loss of bone mass. Maintenance of bone mass showed no significant difference between the OA group and the OVX/DMM+RAL group, but the OVX/DMM+E2 group show significant improvement in bone mass (* P<0.05, ** P<0.01).

Article Snippet: Two months after the models were successfully established, rats were injected with estrogen (0.1mg/kg/day) (Sigma-Aldrich, St. Louis MO, USA) or raloxifene (3mg/kg/day) (MedChem Express, Monmouth Junction, NJ, USA) for one month.

Techniques: Control

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Estrogen Modulates Cartilage and Subchondral Bone Remodeling in an Ovariectomized Rat Model of Postmenopausal Osteoarthritis

doi: 10.12659/MSM.916254

Figure Lengend Snippet: Photomicrographs of the histology of the rat knee joints in the groups treated with estrogen (E2) and raloxifene (RAL). ( A ) The histology of the knee joint in the normal control rat group. Hematoxylin and eosin (H&E) ×100. ( B ) The histology of the knee joint in the ovariectomized/destabilization of the medial meniscus (OVX/DMM) rat group. H&E ×100. ( C ) The histology of the knee joint in the estrogen (E2)-treated rat group. H&E ×100. ( D ) The histology of the knee joint in the raloxifene (RAL)-treated rat group, H&E ×100.

Article Snippet: Two months after the models were successfully established, rats were injected with estrogen (0.1mg/kg/day) (Sigma-Aldrich, St. Louis MO, USA) or raloxifene (3mg/kg/day) (MedChem Express, Monmouth Junction, NJ, USA) for one month.

Techniques: Control

Journal: Scientific reports

Article Title: A Systematic Study of the Impact of Estrogens and Selective Estrogen Receptor Modulators on Prostate Cancer Cell Proliferation.

doi: 10.1038/s41598-020-60844-3

Figure Lengend Snippet: Figure 5. Proliferation of LNCaP cells is increased by E2 only in an AR-dependent manner. LNCaP cell number after treatment, with and without R1881, of either E2 (A), PPT (B), DPN (C), tamoxifen (D), raloxifene (E), bazedoxifene (F), lasofoxifene (G), toremifene (H), and fulvestrant (I). Values were determined with crystal violet staining and are presented as the average ± standard error of the mean (n = 8 samples/treatment). Data from one representative experiment out of six independent experiments is shown. Asterisks (***p < 0.001) indicate that modulation of proliferation is significant in at least four out of six independent experiments.

Article Snippet: The compounds used for treatments were: DMSO (vehicle; Sigma), ethanol 96% (vehicle; Greenfield Global), 17β-estradiol (10 nM; Sigma), PPT (1,3,5-tris(4-hyd roxyphenyl)-4-propyl-1H-pyrazole; Santa Cruz Biotechnology), DPN (2,3-bis(4-hydroxyphenyl) propionitrile; Santa Cruz Biotechnology), fulvestrant (ICI 182,780; Santa Cruz Biotechnology), 4-OH-tamoxifen (Santa Cruz Biotechnology), raloxifene hydrochloride (Tocris), toremifene citrate (Santa Cruz Biotechnology), bazedoxifene (Tocris) and lasofoxifene (Santa Cruz Biotechnology).

Techniques: Staining

Journal: Scientific reports

Article Title: A Systematic Study of the Impact of Estrogens and Selective Estrogen Receptor Modulators on Prostate Cancer Cell Proliferation.

doi: 10.1038/s41598-020-60844-3

Figure Lengend Snippet: Figure 7. Proliferation of 22Rv1 cells is decreased with toremifene (SERM) only when androgens are absent. 22Rv1 cell number after treatment with estrogenic and anti-estrogenic compounds, without (A) and with co-treatment with R1881 (B). Values were determined with crystal violet staining and are presented as the average ± standard error of the mean (n = 8 samples/treatment). The average of two experiments out of five independent experiments is shown. 22Rv1 cell number after treatment with high concentrations of tamoxifen (C) and raloxifene (D) at 10 µM and 28.5 µM, respectively. Data from one representative experiment out of three independent experiments is shown. Asterisks (*p < 0.05, **p < 0.01, ***p < 0.001) indicate that modulation of proliferation is significant compared to controls.

Article Snippet: The compounds used for treatments were: DMSO (vehicle; Sigma), ethanol 96% (vehicle; Greenfield Global), 17β-estradiol (10 nM; Sigma), PPT (1,3,5-tris(4-hyd roxyphenyl)-4-propyl-1H-pyrazole; Santa Cruz Biotechnology), DPN (2,3-bis(4-hydroxyphenyl) propionitrile; Santa Cruz Biotechnology), fulvestrant (ICI 182,780; Santa Cruz Biotechnology), 4-OH-tamoxifen (Santa Cruz Biotechnology), raloxifene hydrochloride (Tocris), toremifene citrate (Santa Cruz Biotechnology), bazedoxifene (Tocris) and lasofoxifene (Santa Cruz Biotechnology).

Techniques: Staining

Journal: Drug Metabolism and Disposition

Article Title: Critical Differences between Enzyme Sources in Sensitivity to Detect Time-Dependent Inactivation of CYP2C8

doi: 10.1124/dmd.118.085498

Figure Lengend Snippet: The initial inactivation constants (kobs) (minute−1) of 100 μM amiodarone (AMD), 100 μM clopidogrel acyl-β-d-glucuronide (CAG), 100 μM gemfibrozil 1-O-β-glucuronide (GFG), 100 μM phenelzine (PHZ), and 1 μM raloxifene (RLX) in different enzyme sources.

Article Snippet: Gemfibrozil 1 -O-β -glucuronide, N -desethylamodiaquine hydrochloride, N -desethylamodiaquine-d5, phenelzine sulfate and raloxifene hydrochloride were from Toronto Research Chemicals (Toronto, ON, Canada).

Techniques:

Journal: bioRxiv

Article Title: Ecological interactions in breast cancer: Cell facilitation promotes growth and survival under drug pressure

doi: 10.1101/2021.02.01.429214

Figure Lengend Snippet: a , Comparing growth of CAMA-1 spheroids under drug pressure with supplemented conditioned media from spheroids of different compositions. Sensitive and resistant cells were plated by the indicated composition of sensitive and resistant cells in a 3D spheroid format and underwent exposure to untreated or 400nM treated medium to produce conditioned media. 100% sensitive spheroids under drug pressure were then supplemented with different concentrations of conditioned media. Normalized cell counts of sensitive cells supplemented with untreated or treated conditioned media generated from different spheroid compositions. b , Liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for estradiol detection performed on CAMA-1 untreated or ribociclib 400nM treated samples of the following compositions at Day 21: 100% sensitive, 50% sensitive – 50% resistant, 100% resistant samples. c , Plots of estradiol production rates of CAMA-1 ribociclib sensitive and resistant cells and sensitive cell uptake of estradiol (relative to resistant cells); estradiol concentration by resistant and sensitive cell abundance d , CAMA-1 growth with modifier treatment in monoculture. Box plots of log of growth for sensitive and resistant cells under indicated modifier treatment (estradiol: p < 0.0001 for interaction between treatment and cell type indicating reduction in growth of resistant cells and increased growth by sensitive cells, tamoxifen: p=0.006 for interaction between treatment and cell type indicating greater reduction in growth of resistant cells, fulvestrant: p=0.011 for interaction between treatment and cell type indicating lower reduction in growth of resistant cells, and raloxifene: no significant interaction with growth reduced equally for the two cells types) e , CAMA-1 growth with combination treatment in monoculture. Comparison of observed cell growth in combination therapy with that expected under null model that assumes no interaction between the effects of the modifier treatment (estradiol, tamoxifen, fulvestrant, or raloxifene) and ribociclib (p < 0.0001 for sensitive cells to grow more rapidly with estradiol and p=0.0007 for sensitive cells to grow more slowly with ER antagonist treatment). f , Box plots of log of growth for sensitive and resistant cells under modifier treatment estradiol or one of three ER antagonists: tamoxifen, fulvestrant, and raloxifene) and ribociclib (p < 0.002 for each ER antagonist to reduce growth of sensitive cells between days 4 and 14 more than resistant cells, but no significant difference among any of the ER antagonists).

Article Snippet: Spheroids were treated with the following drug therapies at specified doses described in results and : ribociclib (Selleck Chemicals, Cat. No: S7440), estradiol (Peprotech, Cat. No: 5022822), fulvestrant (Selleck Chemicals, Cat. No: S1191), tamoxifen (Selleck Chemicals, Cat. No: S7827), and raloxifene (Selleck Chemicals, Cat. No: S5781).

Techniques: Generated, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Concentration Assay, Comparison

Journal: bioRxiv

Article Title: Ecological interactions in breast cancer: Cell facilitation promotes growth and survival under drug pressure

doi: 10.1101/2021.02.01.429214

Figure Lengend Snippet: CAMA-1 spheroids of different cell compositions (100% sensitive - green, 50% sensitive-50% resistant, 100% resistant - blue) cultured in a , untreated, 400nM ribociclib, 0.1nM estradiol, or combination 400nM ribociclib with 0.1nM estradiol treated medium for 18 days; images taken on day 18. b , untreated, 200nM ribociclib, 1 uM tamoxifen, or combination 200nM ribociclib with 1 uM tamoxifen treated medium for 18 days; Images taken on day 18. c , untreated, 400nM ribociclib, 3nM fulvestrant, or combination 400nM ribociclib with 3nM fulvestrant treated medium for 18 days; images taken on day 18. d , untreated, 200nM ribociclib, 75 nM raloxifene, or combination 200nM ribociclib with 75 nM raloxifene treated medium for 18 days; images taken on day 18. e , Facilitation of CAMA-1 growth in coculture measured as the log of the ratio of the observed cell population to the expectation when treated with modifiers (p < 0.0001 for resistant cells to experience greater facilitation than sensitive cells with estradiol, and for sensitive cells to experience greater facilitation than sensitive cells when treated with ER antagonists pooled). The thin dotted lines show facilitation when treated with ribociclib for comparison; Box plots of log of growth for sensitive and resistant cells under treatment (tamoxifen effects not significantly different from estradiol, fulvestrant shows greater facilitation of sensitive cells (p < 0.0001) and raloxifene a smaller effect than fulvestrant (p=0.0035) but greater than estradiol (p=0.0052). f , Facilitation of growth in coculture measured as the log of the ratio of the observed cell population to the expectation when treated with ribociclib in the presence of a modifier (p=0.007 for estradiol to reduce facilitation more for resistant cells, no significant effect with the three ER antagonists pooled). The thin dotted lines show facilitation in the absence of the modifier; Box plots of log of growth for sensitive and resistant cells under combination treatments (no significant differences).

Article Snippet: Spheroids were treated with the following drug therapies at specified doses described in results and : ribociclib (Selleck Chemicals, Cat. No: S7440), estradiol (Peprotech, Cat. No: 5022822), fulvestrant (Selleck Chemicals, Cat. No: S1191), tamoxifen (Selleck Chemicals, Cat. No: S7827), and raloxifene (Selleck Chemicals, Cat. No: S5781).

Techniques: Cell Culture, Comparison

Journal: Journal of separation science

Article Title: Development and validation of ultra-high-performance liquid chromatography–mass spectrometry method for the determination of raloxifene and its phase II metabolites in plasma: Application to pharmacokinetic studies in rats

doi: 10.1002/jssc.202000835

Figure Lengend Snippet: Chemical structures of raloxifene (Ral), raloxifene-6-glucuronide (Ral-6-G), raloxifene-4′-glucuronide (Ral-4′-G), and raloxifene-6-sulfate (Ral-6-S) (A), and the typical chromatograms of Ral(200 nM), Ral-6-G(1000 nM), Ral-4′-G(1000 nM), Ral-6-S(100 nM) and internal standard in LC–MS (B)

Article Snippet: Raloxifene hydrochloride, raloxifene-6-glucuronide, and raloxifene-4′-glucuronide were purchased from Toronto Research Chemicals (Toronto, Canada, all compounds purity ≥99%).

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Cancers

Article Title: New Promising Steroidal Aromatase Inhibitors with Multi-Target Action on Estrogen and Androgen Receptors for Breast Cancer Treatment.

doi: 10.3390/cancers17020165

Figure Lengend Snippet: Figure 6. The impact of ERα on the effects exerted by AIs 6, 10a and 13. (A) MCF-7aro cell viability effects were determined by MTT assay after treatment over 3 days with T (1 nM) plus AIs (1, 5 and 10 µM) in the presence or absence of ICI (100 nM). (B) ER transactivation assays to explore the effects on ER activation, using VM7Luc4E2 cells incubated with AIs (0.1–10 µM), with (ER antagonism) or without (ER agonism) the hormones T or E2. (C) Effects of AIs 6, 10a and 13 (10 µM) on ERα protein expression. β-actin was used as a loading control, being data of densitometry represented as ERα/β-actin ratio. (D) Effects of AIs 6, 10a and 13 (10 µM) on mRNA transcription of ESR1, TFF1, AREG and EGR3 genes in MCF-7aro cells. The mRNA transcript levels of treated cells were quantified using the housekeeping gene ACTB. Cells without treatment were used as control, to which all results in AI-treated cells were normalized. Cells treated with T plus ICI (100 nM) represented positive control. # (p < 0.05), ## (p < 0.01) and ### (p < 0.001) denote differences in MCF-7aro cells treated with AIs in the absence or presence of ICI, while * (p < 0.05), ** (p < 0.01), and *** (p < 0.001) denote differences of AI-treated cells in relation to control cells (T). On the other hand, the differences of AI-treated cells in contrast to control and in relation to ER agonism are indicated by *** (p < 0.001), while in relation to ER antagonism over T are expressed by &&& (p < 0.001).

Article Snippet: For the experiments, VM7Luc4E2 cells were plated at 4 × 105 cells/mL in 96-well white plates and treated with AIs 6, 10a and 13 (0.1–10 μM) over 24 h, with (ER antagonism) or without (ER agonism) 91.8 pM of E2 or 1 nM of T. For agonism assays, we considered as a positive control cells that were treated with T (781.2 pM–25.6 μM) or E2 (180 fM–367 nM), while for ER antagonism, the positive controls were cells that were treated with raloxifene (12.0 pM–24.5 nM; Biosynth Ltd., Berkshire, UK).

Techniques: MTT Assay, Activation Assay, Incubation, Expressing, Control, Positive Control

Journal: Cancers

Article Title: New Promising Steroidal Aromatase Inhibitors with Multi-Target Action on Estrogen and Androgen Receptors for Breast Cancer Treatment.

doi: 10.3390/cancers17020165

Figure Lengend Snippet: Figure 7. The impact of AR on the effects exerted by AIs 6, 10a and 13. (A) MCF-7aro cell viability effects were determined by MTT assay after treatment, over 3 days, with T (1 nM) plus AIs (1, 5 and 10 µM) in the presence or absence of CDX (1 µM). (B) The AR transactivation assay, AR-EcoScreen™, was used to explore the effects of AIs (0.1–10 µM) with (AR antagonism) or without (AR agonism) R1881 (0.1 nM). (C) Effects of AIs (10 µM) on AR protein expression. β-actin was used as a loading control, being data of densitometry represented as AR/β-actin ratio. Cells without AIs treatment were used as control, to which all results in AI-treated cells were normalized. δ (p < 0.05), δδ (p < 0.01) and δδδ (p < 0.001) denote differences of MCF-7aro cells incubated with AIs in the presence or absence of CDX, while ** (p < 0.01) and *** (p < 0.001) denote differences of AI-treated cells in relation to control cells. On the other hand, the differences of AI-treated cells in contrast to control and in relation to AR agonism, are presented by *** (p < 0.001), whereas in relation to AR antagonism are indicated as & (p < 0.05), && (p < 0.001) and &&& (p < 0.001).

Article Snippet: For the experiments, VM7Luc4E2 cells were plated at 4 × 105 cells/mL in 96-well white plates and treated with AIs 6, 10a and 13 (0.1–10 μM) over 24 h, with (ER antagonism) or without (ER agonism) 91.8 pM of E2 or 1 nM of T. For agonism assays, we considered as a positive control cells that were treated with T (781.2 pM–25.6 μM) or E2 (180 fM–367 nM), while for ER antagonism, the positive controls were cells that were treated with raloxifene (12.0 pM–24.5 nM; Biosynth Ltd., Berkshire, UK).

Techniques: MTT Assay, Transactivation Assay, Expressing, Control, Incubation