Journal: Oncology Letters

Article Title: Leptin contributes to the taxol chemoresistance in epithelial ovarian cancer

doi: 10.3892/ol.2019.10381

Figure Lengend Snippet: Leptin reduces the chemosensitivity of ovarian cancer cells to PTX/TXT. The prognosis of patients with epithelial ovarian cancer receiving (A) PTX (P=0.008) or (B) TXT (P=0.0021) chemotherapy was analyzed based on leptin RNA expression level. CCK-8 assay was performed to assess the cell viability of HO8910PM cells treated with DDP combined with (C) PTX/ (D) TXT with or without leptin. CCK-8 assay was performed to assess the HO8910PM cell survival percentage treated with (E) PTX/ (F) TXT with or without leptin. (G) The expression of leptin was highest in the HO8910PM and OV-MZ-15 cell lines. CCK-8 assay was performed to assess the cell viability of OV-MZ-15 cells treated with DDP combined with (H) PTX/ (I) TXT with or without leptin. CCK-8 assay was performed to assess the cell viability of OV-MZ-15 cells treated with (J) PTX/ (K) TXT with or without leptin. Data are presented as the means ± standard deviation. *P<0.05, **P<0.01 and ***P<0.001.

Article Snippet: Recombinant Human Leptin (Bio-Techne) was reconstituted at 1 mg/ml in sterile 20 mM Tris-HCl (pH 8.0) and stored at −80°C.

Techniques: RNA Expression, CCK-8 Assay, Expressing, Standard Deviation

Journal: Oncology Letters

Article Title: Leptin contributes to the taxol chemoresistance in epithelial ovarian cancer

doi: 10.3892/ol.2019.10381

Figure Lengend Snippet: Effects of leptin on the cell cycle distribution of ovarian cancer cells following PTX/TXT treatment. Flow cytometry at (A) 8 and (B) 16 h was used to assess the effect of PTX on the cell cycle distribution of ovarian cancer cells in the presence of leptin. Flow cytometry at (C) 8 and (D) 16 h was used to assess the effect of TXT on the cell cycle distribution of ovarian cancer cells in the presence of leptin. *P<0.05, **P<0.01.

Article Snippet: Recombinant Human Leptin (Bio-Techne) was reconstituted at 1 mg/ml in sterile 20 mM Tris-HCl (pH 8.0) and stored at −80°C.

Techniques: Flow Cytometry

Journal: Oncology Letters

Article Title: Leptin contributes to the taxol chemoresistance in epithelial ovarian cancer

doi: 10.3892/ol.2019.10381

Figure Lengend Snippet: CEBPA initiates the transcription of leptin in ovarian cancer cells. (A) A chromatin immunoprecipitation-polymerase chain reaction assay was performed for HO8910PM and OV-MZ-15 cells. Total DNA (Input), IgG and CEBPA immunoprecipitated DNA were amplified using a series of primers covering the 3,000 bp sequence upstream of the leptin transcription start site. Primer 1 and 9 flanking sequences can be immunoprecipitated with CEBPA. (B) Site directed mutagenesis of the upstream loci of leptin. Leptin vector, leptin-WT or mutant plasmids leptin-MUT1 and 2 were transfected into CEBPA overexpression or control (C) HO8910PM and (D) OV-MZ-15 cells. The pRLTK Renilla luciferase expression plasmid was co-transfected as the internal control. Luciferase activities were normalized to Renilla activities. MUT, mutant; WT, wild-type; CEBPA, CCAAT/enhancer-binding protein α; IgG, immunoglobulin G. **P<0.01, ***P<0.001.

Article Snippet: Recombinant Human Leptin (Bio-Techne) was reconstituted at 1 mg/ml in sterile 20 mM Tris-HCl (pH 8.0) and stored at −80°C.

Techniques: Chromatin Immunoprecipitation, Polymerase Chain Reaction, Immunoprecipitation, Amplification, Sequencing, Mutagenesis, Plasmid Preparation, Transfection, Over Expression, Luciferase, Expressing, Binding Assay

Journal: Oncology Letters

Article Title: Leptin contributes to the taxol chemoresistance in epithelial ovarian cancer

doi: 10.3892/ol.2019.10381

Figure Lengend Snippet: High leptin expression is correlated with a significant enrichment of multiple hallmark of cancer gene sets. (A) A total of 46 patients with ovarian cancer treated with PTX were assessed and divided into two groups according to leptin expression. ***P<0.001. (B) A heatmap of differentially expressed genes between groups associated with low leptin expression one and two was constructed group 1, leptin low expression; group 2, leptin high expression; ‘red’ means upregulation and ‘green’ means downregulation). (C) Results from the gene set enrichment analysis revealed the leptin-associated enrichment gene sets with the highest NES value. (D-I) The test leptin-associated enrichment gene sets were closely associated with tumor progression and chemoresistance and included the following hallmark: (D) HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITION; (E) HALLMARK_TNFA_SIGNALING_VIA_NFKB; (F) HALLMARK_KRAS_SIGNALING_UP; (G) HALLMARK_HYPOXIA; (H) KEGG_ECM_RECEPTOR_INTERACTION; and (I) HALLMARK_IL6_JAK_STAT3_SIGNALING NES, Normalized Enrichment Score.

Article Snippet: Recombinant Human Leptin (Bio-Techne) was reconstituted at 1 mg/ml in sterile 20 mM Tris-HCl (pH 8.0) and stored at −80°C.

Techniques: Expressing, Construct

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) MALDI-TOF spectra of reaction mixture. The ratio denotes the molar ratio of leptin monomer to polymer. (b) SEC of leptin conjugates eluted from Superdex75 100/300 GL column in 10% MeOH/0.25 M sodium phosphate, pH 7.5 at 0.5 ml/min. (c) SDS-PAGE of pooled fractions in SEC denoted by elution time. Fraction of 20.3–21.9 min in LepNP85 and fraction of 18.5–21 min in LepNPEG5K were used.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Polymer, SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: SDS-PAGE of the reaction mixture of leptin and Leptin-2PCA with CHO-P85-OH or with P85-DSP.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: SDS Page

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Leptin 1:1 physical mixture with P85 or PEG5K. (b) Leptin 1:1 conjugates.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Leptin and leptin 1:1 conjugates were eluted from Jupiter C4 column (particle diameter 5 μm, pore diameter 300 Å, 4.6 × 100 mm) by gradient elution at 1 ml/min and 25 °C, and monitored by absorption at 220 nm. Mobile phase A: water + 0.1% TFA; mobile phase B: acetonitrile + 0.1% TFA: isopropanol 50:50 (v/v). The elution started from 5% B for 5 min, then linearly increased to 95% B at 1%/min, and stayed at 95% B till 100 min.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Representative sensorgram for native leptin. (b) Representative sensorgram for LepNP85 1:1 conjugates. (c) Representative sensorgram for LepDSSP85 1:1 conjugates. (d) Dissociation constants (KD) of leptin and leptin 1:1 conjugates. Data are mean ± SD, n= 5~12. *** p < 0.001 and n.s. not significant by One-way ANOVA and post Newman-Keuls Multiple Comparison Test.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Comparison

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: (a) Dose response of leptin and LepNP85 1:1 conjugates. Data are mean ± SEM, n=3. (b) Comparison of leptin and leptin 1:1 conjugates at 100 ng/mouse. Data are mean ± SEM, n= 5~7. ** p < 0.01 and *** p < 0.001 by One-way ANOVA and post Newman-Keuls Multiple Comparison Test.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Comparison

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Serum clearance, unidirectional brain influx rates and initial volumes of brain distribution for leptin and leptin 1:1 conjugates.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Male CD-1 mice were co-injected with 125I-LepNP85 and 131I-leptin. Data are mean ± SEM, n=7/time point, * p < 0.05, ** p < 0.01, and *** p < 0.001 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: Each male CD-1 mouse was injected with 125I-labeled LepNP85 with or without 10 μg of cold leptin. Data are mean ± SEM, n=7/time point, * p < 0.05 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection, Labeling

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Intranasal Delivery of N-terminal Modified Leptin-Pluronic Conjugate for Treatment of Obesity

doi: 10.1016/j.jconrel.2017.03.029

Figure Lengend Snippet: 1 μg of biotin-labeled leptin and LepNP85 in 1 μl of PBS were injected locally into brain by ICV injection. The concentrations of biotin-labeled proteins in brain lysate and plasma were analyzed by ELISA. (a) Blood absorption of biotin-leptin and biotin-LepNP85 after ICV injection. (b) Brain retention of biotin-leptin and biotin-LepNP85 after ICV injection. Data are mean ± SEM, n=4/time point, * p < 0.05 by One-way ANOVA and post Dunnett’s Multiple Comparison Test, and *** p < 0.001 by Two-way ANOVA.

Article Snippet: Recombinant mouse leptin and recombinant human leptin receptor-Fc chimeras were purchased from R&D Systems (Minneapolis, MN).

Techniques: Labeling, Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Indian Journal of Endocrinology and Metabolism

Article Title: Leptin therapy, insulin sensitivity, and glucose homeostasis

doi: 10.4103/2230-8210.105571

Figure Lengend Snippet: Euglycemic hyperinsulinemic clamps of leptin-deficient patients while on leptin and after brief periods of leptin withdrawal (from Paz-Filho et al .). Glucose infusion rates increased in eight out of nine times when leptin therapy was briefly interrupted. The substantial weight gain after leptin withdrawal was responsible for acutely increasing insulin sensitivity

Article Snippet: [ ] Physiological doses of recombinant methionyl human leptin (r-metHuLeptin, Metreleptin ® , Amylin Pharmaceuticals, USA, 0.02-0.04 mg/kg/day) were initiated on the leptin-deficient individuals being assessed by our group at ages 5 (boy), 27 (male), 30 and 40 (females).

Techniques:

Journal: Indian Journal of Endocrinology and Metabolism

Article Title: Leptin therapy, insulin sensitivity, and glucose homeostasis

doi: 10.4103/2230-8210.105571

Figure Lengend Snippet: Interactions between fat, brain, pancreas and bone. Leptin activates the sympathetic tonus, which inhibits insulin secretion and stimulates Esp expression in the osteoblast (via stimulation of the adrenergic beta 2 receptor). Esp inhibits osteocalcin, decreasing insulin expression. Leptin inhibits pancreatic insulin. Insulin, in turn, stimulates leptin expression in the white adipose tissue. Adapted from Kieffer and Habener. H: Hypothalamus

Article Snippet: [ ] Physiological doses of recombinant methionyl human leptin (r-metHuLeptin, Metreleptin ® , Amylin Pharmaceuticals, USA, 0.02-0.04 mg/kg/day) were initiated on the leptin-deficient individuals being assessed by our group at ages 5 (boy), 27 (male), 30 and 40 (females).

Techniques: Expressing