allo aca  (MedChemExpress)

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: Leptin upregulates LEPR expression in TNBC cells. (A, B) Bioinformatics data from Kaplan-Meier database ( https://kmplot.com/analysis/index.php?p=service ) showed OS curve (A) and DMFS curve (B) among TNBC patients stratified by high and low LEPR expression levels. Higher LEPR expression was associated with poor prognosis. (C, D) Quantitative PCR and western blot were performed to analyze the expression of LEPR mRNA (C) and LEPR protein (D) in 4T1 and EMT6 cells following stimulation with increasing concentrations of leptin (0, 10, 50, 100, 200 ng/mL) for 24 hours, respectively. n=3-4. (E, F) Quantitative LEPR mRNA (E) and LEPR protein (F) expression in 4T1 and EMT6 cells following 24 hours of leptin (100 ng/mL) and/or Allo-aca (Allo, 100 nM) treatment. n=3. DMFS, distant metastases-free survival; HR, hazard ratio; LEPR, leptin receptor; OS, overall survival; PCR, polymerase chain reaction; SEM, standard error of mean; TNBC, triple-negative breast cancer. All data are presented as the mean±SEM. ns, no significance; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Additionally, the LEPR antagonist, Allo-aca (100 nM for 24 hours, MCE), and the specific ADRB2 inhibitor, ICI118551 (10 μM for 24 hours, Sigma-Aldrich), were separately added to 4T1 and EMT6 cells at appropriate densities prior to further experimentation.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Polymerase Chain Reaction

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: Leptin upregulates ADRB2 expression in TNBC cells. (A) Quantitative PCR was utilized to assess the expression levels of ADRB2 in 4T1 (n=3) and EMT6 (n=6) cells following stimulation with 50 ng/mL or 100 ng/mL leptin. (B) Western blotting was utilized to assess the expression levels of ADRB2 in 4T1 and EMT6 cells following stimulation with 50 ng/mL or 100 ng/mL leptin. n=4. (C, D) Quantitative PCR (C) and western blotting (D) were employed to analyze the expression of ADRB2 in 4T1 and EMT6 cells treated with leptin (100 ng/mL) and/or Allo (100 nM). n=3. (E) Representative images of immunofluorescence double staining showing the expression and co-localization of LEPR (Alexa Fluor 488 Conjugated) and ADRB2 (Alexa Fluor 594-conjugated) in the presence of leptin and/or Allo, or the ADRB2 inhibitor ICI118551 (ICI). (F-J) Statistical curves depicting the fluorescence intensity of LEPR and ADRB2 in the co-localization images for different experimental groups. (K) Quantitative analysis of LEPR-ADRB2 co-localization on cell membranes in each group, evaluated using Pearson’s correlation coefficient. Data are presented as mean±SEM, n=3. ADRB2, adrenergic receptor β2; LEPR, leptin receptor; PCR, polymerase chain reaction; SEM, standard error of mean; TNBC, triple-negative breast cancer. All data are presented as the mean±SEM. ns, no significance; * p < 0.05, ** p < 0.01.

Article Snippet: Additionally, the LEPR antagonist, Allo-aca (100 nM for 24 hours, MCE), and the specific ADRB2 inhibitor, ICI118551 (10 μM for 24 hours, Sigma-Aldrich), were separately added to 4T1 and EMT6 cells at appropriate densities prior to further experimentation.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Double Staining, Fluorescence, Polymerase Chain Reaction

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: LEPR and ADRB2 in leptin-induced breast cancer progression in vivo . (A) Female BALB/c mice were subcutaneously inoculated with 1×10 5 4T1 cells into the mammary fat pad to establish the breast tumor animal model. Drug administration commenced 7 days postinoculation and continued for 21 days. Tumors, blood samples, and lungs were collected for analysis. (B) Representative image of tumors and tumor growth curves in different experimental groups. n=5. (C) Tumor weights were measured on day 28. n=5. (D, E) Food intake (D) and body weight curves (E) in different groups. n=5. (F) Metastatic lung nodules were visualized using H&E staining in different groups. (G) GSH content was assessed in both serum and tumor samples collected from different groups. n=3-5. (H, I) Real-time qPCR (H) and western blotting (I) were used to analyze the expression of LEPR, ADRB2, and NOX4 in tumors from different groups. n=3. ADRB2, adrenergic receptor β2; GSH, glutathione; LEPR, leptin receptor; NOX4, NADPH oxidase 4; qPCR, quantitative real-time polymerase chain reacion; SEM, standard error of mean. All data are presented as the mean±SEM. ns, no significance; * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Additionally, the LEPR antagonist, Allo-aca (100 nM for 24 hours, MCE), and the specific ADRB2 inhibitor, ICI118551 (10 μM for 24 hours, Sigma-Aldrich), were separately added to 4T1 and EMT6 cells at appropriate densities prior to further experimentation.

Techniques: In Vivo, Animal Model, Staining, Western Blot, Expressing

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Synergistic Activation of LEPR and ADRB2 Induced by Leptin Enhances Reactive Oxygen Specie Generation in Triple-Negative Breast Cancer Cells

doi: 10.4143/crt.2024.368

Figure Lengend Snippet: The clinical significance of LEPR and ADRB2. (A, B) Receiver operating characteristic curve analysis was conducted to evaluate the prognostic significance of LEPR (A) and ADRB2 (B) in breast cancer patients. p < 0.01. (C-E) The correlation between LEPR, ADRB2, NOX4, and GPX4 was analyzed using the bc-GenExMiner database (C) and GEPIA database (D, E). (F) Kaplan-Meier plotter was utilized to illustrate RFS in TNBC patients with high expression of LEPR and ADRB2 combined. p=0.0071. (G) The schematic diagram of the study work. ADRB2, adrenergic receptor β2; AUC, area under the curve; GPX4, glutathione peroxidase 4; HR, hazard ratio; LEPR, leptin receptor; mtROS, mitochondrial reactive oxygen species; NOX4, NADPH oxidase 4; RFS, relapse-free survival; TNBC, triple-negative breast cancer; TNR, true-negative ratio; TPM, transcripts per million; TPR, true-positive ratio.

Article Snippet: Additionally, the LEPR antagonist, Allo-aca (100 nM for 24 hours, MCE), and the specific ADRB2 inhibitor, ICI118551 (10 μM for 24 hours, Sigma-Aldrich), were separately added to 4T1 and EMT6 cells at appropriate densities prior to further experimentation.

Techniques: Expressing