Journal: Pharmacology Research & Perspectives

Article Title: Challenges of measuring accurate estradiol levels in aromatase inhibitor‐treated postmenopausal breast cancer patients on vaginal estrogen therapy

doi: 10.1002/prp2.330

Figure Lengend Snippet: Comparison of estradiol levels in breast cancer patients on AI being treated with vaginal estradiol by ELISA versus estimation by LC ‐ MS / MS method: The serum samples were collected at various intervals from three patients who were on Estring ® to measure estradiol concentrations by two different ELISA kits, Invitrogen, and IBL . A few of these samples were also submitted for LC ‐ MS / MS analysis conducted by ARUP laboratories for the comparison purposes. The red line indicates limit of detection (LOD) for ELISA. 2A. Patient #1 was on letrozole and it was found that estradiol levels were varied between two different ELISA kits. 2B. The estrogen levels measured for patient #5 showed similar results by two ELISA kits at baseline, week 2 and week 4 (<3.0 pg/dl); whereas for week 8, the levels by Invitrogen kit was higher than IBL kit. Discrepancy was also found in week 2 results (8.6 pg/mL) by LC ‐ MS / MS analysis. 2C. The baseline estradiol results for patient #7 who was taking exemestane were highest at baseline determined by both IBL kit and LC ‐ MS / MS assay. 2D. Retesting of a few stored samples was done and consistency in the estradiol values was observed.

Article Snippet: This suggests that the IBL kit may have had more cross‐reactivity with exemestane than the Invitrogen kit.

Techniques: Comparison, Enzyme-linked Immunosorbent Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Pharmacology Research & Perspectives

Article Title: Challenges of measuring accurate estradiol levels in aromatase inhibitor‐treated postmenopausal breast cancer patients on vaginal estrogen therapy

doi: 10.1002/prp2.330

Figure Lengend Snippet: Patient demographics with history of stage I–III estrogen receptor‐positive breast cancer taking adjuvant aromatase inhibitor therapy

Article Snippet: This suggests that the IBL kit may have had more cross‐reactivity with exemestane than the Invitrogen kit.

Techniques: Adjuvant

Journal: Molecules

Article Title: In Vitro Effects of Combining Genistein with Aromatase Inhibitors: Concerns Regarding Its Consumption during Breast Cancer Treatment

doi: 10.3390/molecules28134893

Figure Lengend Snippet: Effects of the combination of Genistein (G) with AIs Exemestane (Exe) ( A ), Anastrozole (Ana) ( B ), and Letrozole (Let) ( C ) on the viability of MCF-7aro cells. Cells stimulated with testosterone (T; 1 nM) were treated with G (1 or 10 µM) and Exe (1, 5 or 10 µM), Ana (1, 5 or 10 µM), or Let (1, 5 or 10 µM) for 3 days. Cells only treated with T were considered the control group (100% of cell viability). Data are presented as mean ± SEM of at least three independent experiments, performed in triplicate. Significant differences between control and treated cells are designated as * ( p < 0.05), ** ( p < 0.01), *** ( p < 0.001), and **** ( p < 0.0001), while differences between the AIs and the combination of G with AIs are indicated as # ( p < 0.05), ## ( p < 0.01) and ### ( p < 0.001).

Article Snippet: Exemestane (Exe) (Carbosynth, Berkshire, UK), Genistein (G) from Glycine max , Anastrazole (Ana), Letrozole (Let), Casodex (CDX), ICI 182,780 and 4-[2-phenyl-5,7, bis (trifluoromethyl) pyrazole [1,5-a]pyrimidin-3-yl]phenol (PHTPP) (Sigma-Aldrich Co., Saint Louis, MO, USA) were prepared in 100% DMSO (Sigma-Aldrich Co., Saint Louis, MO, USA).

Techniques: Control

Journal: Molecules

Article Title: In Vitro Effects of Combining Genistein with Aromatase Inhibitors: Concerns Regarding Its Consumption during Breast Cancer Treatment

doi: 10.3390/molecules28134893

Figure Lengend Snippet: Effects on cell cycle of MCF-7aro of Genistein (G) alone or in combination with Exemestane (Exe) for 3 days.

Article Snippet: Exemestane (Exe) (Carbosynth, Berkshire, UK), Genistein (G) from Glycine max , Anastrazole (Ana), Letrozole (Let), Casodex (CDX), ICI 182,780 and 4-[2-phenyl-5,7, bis (trifluoromethyl) pyrazole [1,5-a]pyrimidin-3-yl]phenol (PHTPP) (Sigma-Aldrich Co., Saint Louis, MO, USA) were prepared in 100% DMSO (Sigma-Aldrich Co., Saint Louis, MO, USA).

Techniques: Control

Journal: Molecules

Article Title: In Vitro Effects of Combining Genistein with Aromatase Inhibitors: Concerns Regarding Its Consumption during Breast Cancer Treatment

doi: 10.3390/molecules28134893

Figure Lengend Snippet: Effects of the combination of Genistein (G) with Exemestane (Exe) on the expression levels of cyclin E, cleaved PARP, and pro-caspase-7 in MCF-7aro cells. Cells were stimulated with Testosterone (T) (1 nM) and treated with G (1 or 10 µM) with or without Exe (10 µM) for 3 days. Cells treated only with T were considered the control group. Representative Western blot ( A ) and densitometric analysis ( B ) of cyclin E, cleaved PARP, and pro-caspase-7. β-Actin or total PARP was used as a loading control. The protein expression obtained for treated cells was normalized in relation to the protein expression of the control. The results are presented as mean ± SEM of at least three independent experiments. Significant differences between control and treated cells are designated as * ( p < 0.05), ** ( p < 0.01), *** ( p < 0.001) and **** ( p < 0.0001), while differences between Exe and its combination with G are indicated as # ( p < 0.05) and ## ( p < 0.01).

Article Snippet: Exemestane (Exe) (Carbosynth, Berkshire, UK), Genistein (G) from Glycine max , Anastrazole (Ana), Letrozole (Let), Casodex (CDX), ICI 182,780 and 4-[2-phenyl-5,7, bis (trifluoromethyl) pyrazole [1,5-a]pyrimidin-3-yl]phenol (PHTPP) (Sigma-Aldrich Co., Saint Louis, MO, USA) were prepared in 100% DMSO (Sigma-Aldrich Co., Saint Louis, MO, USA).

Techniques: Expressing, Control, Western Blot

Journal: Molecules

Article Title: In Vitro Effects of Combining Genistein with Aromatase Inhibitors: Concerns Regarding Its Consumption during Breast Cancer Treatment

doi: 10.3390/molecules28134893

Figure Lengend Snippet: Influence of aromatase on the effects induced by Genistein (G) alone and when combined with Exemestane (Exe) in MCF-7aro cells. ( A ) Effects of G (1–10 µM) on the viability of MCF-7aro cells stimulated with Testosterone (T) (1 nM) or estradiol (E 2 ) (1 nM), after 3 days. Cells treated only with T or E 2 were considered the control group. ( B ) Representative Western blot and densitometric analysis of the effects of G (1 and 10 µM) with or without Exe (10 µM) on the expression levels of aromatase, after 8 h exposure. Β-Actin was used as a loading control. The protein expression obtained for treated cells was normalized in relation to the protein expression of the control (T-treated cells). The results are presented as mean ± SEM of at least three independent experiments. Significant differences between control and treated cells are designated as ** ( p < 0.01) and *** ( p < 0.001) while differences between cells treated with G and stimulated with T versus E 2 are denoted as & ( p < 0.05) and &&&& ( p < 0.0001), and differences between Exe and its combination with G are indicated as ## ( p < 0.01). MIC: placenta microsomes (positive control).

Article Snippet: Exemestane (Exe) (Carbosynth, Berkshire, UK), Genistein (G) from Glycine max , Anastrazole (Ana), Letrozole (Let), Casodex (CDX), ICI 182,780 and 4-[2-phenyl-5,7, bis (trifluoromethyl) pyrazole [1,5-a]pyrimidin-3-yl]phenol (PHTPP) (Sigma-Aldrich Co., Saint Louis, MO, USA) were prepared in 100% DMSO (Sigma-Aldrich Co., Saint Louis, MO, USA).

Techniques: Control, Western Blot, Expressing, Positive Control

Journal: Molecules

Article Title: In Vitro Effects of Combining Genistein with Aromatase Inhibitors: Concerns Regarding Its Consumption during Breast Cancer Treatment

doi: 10.3390/molecules28134893

Figure Lengend Snippet: Influence of hormone receptors ERα, ERβ, and AR on the effects induced by Genistein (G) alone and when combined with Exemestane (Exe) in MCF-7aro cells. ( A ) Effects of G (1–10 µM) on the viability of MCF-7aro cells stimulated with Testosterone (T) (1 nM) and treated with or without ICI (100 nM) or PHTPP (1 µM) or CDX (1 µM), after 3 days. Cells treated only with T with or without ICI, PHTPP, or CDX were considered the control group. ( B ) Representative Western blot and densitometric analysis of the effects of G (1 and 10 µM) with or without Exe (10 µM) on the expression levels of ERα, ERβ, and AR, after 3 days. β-Actin was used as a loading control. The protein expression obtained for treated cells was normalized in relation to the protein expression of the control (T-treated cells). The results are presented as mean ± SEM of at least three inde-pendent experiments. Significant differences between control and treated cells are designated as ** ( p < 0.01), *** ( p < 0.001), and **** ( p < 0.0001), while differences between cells treated with G and incubated with T with or without ICI, PHTPP or CDX are denoted as & ( p < 0.05) and &&&& ( p < 0.0001).

Article Snippet: Exemestane (Exe) (Carbosynth, Berkshire, UK), Genistein (G) from Glycine max , Anastrazole (Ana), Letrozole (Let), Casodex (CDX), ICI 182,780 and 4-[2-phenyl-5,7, bis (trifluoromethyl) pyrazole [1,5-a]pyrimidin-3-yl]phenol (PHTPP) (Sigma-Aldrich Co., Saint Louis, MO, USA) were prepared in 100% DMSO (Sigma-Aldrich Co., Saint Louis, MO, USA).

Techniques: Control, Western Blot, Expressing, Incubation