Journal: Annals of Medicine

Article Title: Human adipose-derived stem cells promote migration of papillary thyroid cancer cell via leptin pathway

doi: 10.1080/07853890.2024.2419990

Figure Lengend Snippet: Leptin receptor (Ob-R) antagonist reduced the number of metastatic lung nodules and the diameter of nodules. (A) Procedure of animal experiment. Nude mice received venous transplantation of TPC-1 cells. To block OB-R, 1 mg/kg of Allo-aca was administrated to the animal per day. (B) Gross morphology of mice lungs showed nodules. Allo-aca reduced the number of pulmonary nodules. (C) Lung nodules were diagnosed as metastatic tumour under the light microscope. Allo-aca reduced the diameter of the nodules. (D) The lung nodules were examined by Western blot of thyroglobulin (TG). The nodules in the lungs were TG -positive, while the lung tissue was TG -negative. This result confirmed that the lung nodules were from thyroid carcinoma. (E–H) Allo-aca increased the level of serum T3, and had no significant effect on the level of T4, TSH and the body weight of the animal. Experiments were repeated at least five times. Data are expressed as mean ± standard deviation. * p < 0.05, *** p < 0.001, ns: not statistically significant.

Article Snippet: The PVDF membranes were blocked with BSA (Sigma-Aldrich) and then hybridized with antibodies, including the following: mouse monoclonal IgG of leptin receptor (Ob-R) antibody (R&D Systems, catalog# MAB867, diluted 1:500), rabbit polyclonal MMP-2 (Cell Signaling Technology, catalog# 4022, diluted 1:1000), MMP-9 antibodies (Signaling Technology, catalog# 3852, diluted 1:1000), rabbit monoclonal thyroglobulin antibody (Abcam, catalog# ab243094, diluted 1:1000), rabbit polyclonal GAPDH antibody (Abcam, catalog# ab9485, diluted 1:1000), goat anti-mouse IgG (Abcam, catalog# ab6789, diluted 1:2000), goat anti-rabbit IgG (Abcam, catalog# ab205718, diluted 1:2000).

Techniques: Transplantation Assay, Blocking Assay, Light Microscopy, Western Blot, Standard Deviation

Journal: Annals of Medicine

Article Title: Human adipose-derived stem cells promote migration of papillary thyroid cancer cell via leptin pathway

doi: 10.1080/07853890.2024.2419990

Figure Lengend Snippet: The conditioned medium of ADSCs upregulated the level of leptin receptor (Ob-R) in TPC-1 and BCPAP cells. (A) The concentration of leptin in the medium of ADSCs was ∼3-fold higher than that in the medium of TPC-1 and BCPAP cells, approximately 50 pg/ml. (B) In vitro experiments showed that ADSC-CM increased the level of leptin receptor (Ob-R) in the TPC-1/BCPAP cells. Experiments were repeated at least five times. Data are expressed as mean ± standard deviation. * p < 0.05, *** p < 0.001.

Article Snippet: The PVDF membranes were blocked with BSA (Sigma-Aldrich) and then hybridized with antibodies, including the following: mouse monoclonal IgG of leptin receptor (Ob-R) antibody (R&D Systems, catalog# MAB867, diluted 1:500), rabbit polyclonal MMP-2 (Cell Signaling Technology, catalog# 4022, diluted 1:1000), MMP-9 antibodies (Signaling Technology, catalog# 3852, diluted 1:1000), rabbit monoclonal thyroglobulin antibody (Abcam, catalog# ab243094, diluted 1:1000), rabbit polyclonal GAPDH antibody (Abcam, catalog# ab9485, diluted 1:1000), goat anti-mouse IgG (Abcam, catalog# ab6789, diluted 1:2000), goat anti-rabbit IgG (Abcam, catalog# ab205718, diluted 1:2000).

Techniques: Concentration Assay, In Vitro, Standard Deviation

Journal: Annals of Medicine

Article Title: Human adipose-derived stem cells promote migration of papillary thyroid cancer cell via leptin pathway

doi: 10.1080/07853890.2024.2419990

Figure Lengend Snippet: The neutralizing antibody to leptin reduced the invasion and migration of the TPC-1 and BCPAP cells. ADSC-CM upregulated MMP-2 in TPC-1 and BCPAP cells, and the effect was attenuated by the neutralizing antibody to leptin. (A) Addition of 10 μg/mL NAB of leptin to ADSC-CM conditioned medium attenuated TPC-1 and BCPAP invasion. (B) Leptin NAB also attenuated conditioned medium-promoted TPC-1/BCPAP migration. Experiments were repeated at least five times. Data are expressed as mean ± standard deviation. * p < 0.05, *** p < 0.001.

Article Snippet: The PVDF membranes were blocked with BSA (Sigma-Aldrich) and then hybridized with antibodies, including the following: mouse monoclonal IgG of leptin receptor (Ob-R) antibody (R&D Systems, catalog# MAB867, diluted 1:500), rabbit polyclonal MMP-2 (Cell Signaling Technology, catalog# 4022, diluted 1:1000), MMP-9 antibodies (Signaling Technology, catalog# 3852, diluted 1:1000), rabbit monoclonal thyroglobulin antibody (Abcam, catalog# ab243094, diluted 1:1000), rabbit polyclonal GAPDH antibody (Abcam, catalog# ab9485, diluted 1:1000), goat anti-mouse IgG (Abcam, catalog# ab6789, diluted 1:2000), goat anti-rabbit IgG (Abcam, catalog# ab205718, diluted 1:2000).

Techniques: Migration, Standard Deviation

Journal: Annals of Medicine

Article Title: Human adipose-derived stem cells promote migration of papillary thyroid cancer cell via leptin pathway

doi: 10.1080/07853890.2024.2419990

Figure Lengend Snippet: Neutralizing antibodies against leptin slightly downregulated cell proliferation without significant effect on cell viability. (A) Addition of leptin NAB to ADSC-CM slightly reduced the stimulatory effect of conditioned medium on TPC-1 proliferation. (B) Addition of NAB to the conditioned medium had no significant effect on cell viability of TPC-1 cells. (C) NAB of leptin had no significant effect on BCPAP proliferation. (D) NAB of leptin had no significant effect on BCPAP viability. Experiments were repeated at least five times. Data are expressed as mean ± standard deviation. * p < 0.05, ns: not statistically significant.

Article Snippet: The PVDF membranes were blocked with BSA (Sigma-Aldrich) and then hybridized with antibodies, including the following: mouse monoclonal IgG of leptin receptor (Ob-R) antibody (R&D Systems, catalog# MAB867, diluted 1:500), rabbit polyclonal MMP-2 (Cell Signaling Technology, catalog# 4022, diluted 1:1000), MMP-9 antibodies (Signaling Technology, catalog# 3852, diluted 1:1000), rabbit monoclonal thyroglobulin antibody (Abcam, catalog# ab243094, diluted 1:1000), rabbit polyclonal GAPDH antibody (Abcam, catalog# ab9485, diluted 1:1000), goat anti-mouse IgG (Abcam, catalog# ab6789, diluted 1:2000), goat anti-rabbit IgG (Abcam, catalog# ab205718, diluted 1:2000).

Techniques: Standard Deviation

Journal: Annals of Medicine

Article Title: Human adipose-derived stem cells promote migration of papillary thyroid cancer cell via leptin pathway

doi: 10.1080/07853890.2024.2419990

Figure Lengend Snippet: (A,B) The ADSCs conditioned medium upregulated the MMP-2 levels of TPC-1/BCPAP. NAB of leptin partially counteracted the increase in MMP-2 levels. Experiments were repeated at least five times. Data are expressed as mean ± standard deviation. * p < 0.05, ** p < 0.01.

Article Snippet: The PVDF membranes were blocked with BSA (Sigma-Aldrich) and then hybridized with antibodies, including the following: mouse monoclonal IgG of leptin receptor (Ob-R) antibody (R&D Systems, catalog# MAB867, diluted 1:500), rabbit polyclonal MMP-2 (Cell Signaling Technology, catalog# 4022, diluted 1:1000), MMP-9 antibodies (Signaling Technology, catalog# 3852, diluted 1:1000), rabbit monoclonal thyroglobulin antibody (Abcam, catalog# ab243094, diluted 1:1000), rabbit polyclonal GAPDH antibody (Abcam, catalog# ab9485, diluted 1:1000), goat anti-mouse IgG (Abcam, catalog# ab6789, diluted 1:2000), goat anti-rabbit IgG (Abcam, catalog# ab205718, diluted 1:2000).

Techniques: Standard Deviation

Journal: Annals of Medicine

Article Title: Human adipose-derived stem cells promote migration of papillary thyroid cancer cell via leptin pathway

doi: 10.1080/07853890.2024.2419990

Figure Lengend Snippet: Hypothesis diagram. Adipose tissue acts on thyroid cancer cells by secreting leptin. Leptin upregulates the level of the leptin receptor (Ob-R) on the surface of thyroid cancer cells and up-modulates the level of MMP-2 by activating the leptin signalling pathway, which may be an important mechanism by which obesity promotes the metastasis of thyroid cancer cells.

Article Snippet: The PVDF membranes were blocked with BSA (Sigma-Aldrich) and then hybridized with antibodies, including the following: mouse monoclonal IgG of leptin receptor (Ob-R) antibody (R&D Systems, catalog# MAB867, diluted 1:500), rabbit polyclonal MMP-2 (Cell Signaling Technology, catalog# 4022, diluted 1:1000), MMP-9 antibodies (Signaling Technology, catalog# 3852, diluted 1:1000), rabbit monoclonal thyroglobulin antibody (Abcam, catalog# ab243094, diluted 1:1000), rabbit polyclonal GAPDH antibody (Abcam, catalog# ab9485, diluted 1:1000), goat anti-mouse IgG (Abcam, catalog# ab6789, diluted 1:2000), goat anti-rabbit IgG (Abcam, catalog# ab205718, diluted 1:2000).

Techniques:

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Estradiol affects extracellular leptin:adiponectin ratio in human breast tissue in vivo.

doi: 10.1210/jc.2014-1129

Figure Lengend Snippet: Figure 3. Tamoxifen significantly decreased leptin and increased adiponectin in normal human breast tissue. A. Postmenopausal women underwent microdialysis sampling of breast tissue (n 18) and subcutaneous (sc) abdominal fat (n 16) before tamoxifen treatment and 6 weeks after treatment. *P .05, **P .01. B. To confirm the in vivo findings, whole normal human breast tissue biopsies were cultured with or without 10-6 M tamoxifen (Tam) for 7 days, and adipokine levels in the culture medium were determined. *P .05 (n 9), ****P .0001 (n 12) compared to controls. C. Normal breast tissue biopsies were cultured ex vivo with or without 10-6 M tamoxifen (Tam). Levels of adipokines and their corresponding receptors (leptin receptor [LepR], adiponectin receptor 1 [AdipoR1], and adiponectin receptor 2 [AdipoR2]) were determined by immunohistochemistry analysis. No differences were detected between treatment groups. Representative tissue sections from each treatment group are shown (scale bars, 20 m).

Article Snippet: Immunohistochemistry Formalin-fixed, paraffin-embedded normal breast tissue biopsies and mouse tumors were cut into 4- m sections, deparaffinized, and subjected to high-temperature antigen retrieval prior to incubating with the following primary antibodies: rabbit antihuman leptin (1 g/ml; GenWay Biotech, San Diego, CA, USA), goat antihuman LepR (20 g/ml; AbD Serotec, Düsseldorf, Germany), goat antimouse leptin (10 g/ml; R&D Systems), goat antimouse LepR (10 g/ml; R&D Systems), rabbit antihuman adiponectin (1 g/ml; Abcam, Cambridge, UK), rabbit antihuman AdipoR1 (5 g/ml; Bioss, London, UK), and rabbit antihuman AdipoR2 (5 g/ml; Bioss).

Techniques: Sampling, In Vivo, Cell Culture, Ex Vivo, Immunohistochemistry

Journal: The Journal of clinical endocrinology and metabolism

Article Title: Estradiol affects extracellular leptin:adiponectin ratio in human breast tissue in vivo.

doi: 10.1210/jc.2014-1129

Figure Lengend Snippet: Figure 4. Increased extracellular leptin and decreased adiponectin in human breast cancer in vivo. A. Eleven postmenopausal breast cancer patients underwent microdialysis sampling of the tumor and normal adjacent breast tissue before surgery. *P .05, **P .01. B. FVB/N mice were oophorectomized and treated with or without estradiol (physiologic level), and MMTV- PyMT mammary cancer cells were injected into the dorsal mammary fat pad. At similar tumor sizes, both groups of mice underwent microdialysis sampling of adipokines in the tumor. *P .05, **P .01 (each group, n 7). C. FVB/N mice were treated as described in Figure 4B. Levels of adipokines and their corresponding receptors (leptin receptor [LepR], adiponectin receptor 1 [AdipoR1], and adiponectin receptor 2 [AdipoR2]) in the MMTV-PyMT tumors were determined by immunohistochemistry analysis. Increased LepR staining was observed in estrogen-exposed tumors. Representative tissue sections from each treatment group are shown (scale bars, 20 m).

Article Snippet: Immunohistochemistry Formalin-fixed, paraffin-embedded normal breast tissue biopsies and mouse tumors were cut into 4- m sections, deparaffinized, and subjected to high-temperature antigen retrieval prior to incubating with the following primary antibodies: rabbit antihuman leptin (1 g/ml; GenWay Biotech, San Diego, CA, USA), goat antihuman LepR (20 g/ml; AbD Serotec, Düsseldorf, Germany), goat antimouse leptin (10 g/ml; R&D Systems), goat antimouse LepR (10 g/ml; R&D Systems), rabbit antihuman adiponectin (1 g/ml; Abcam, Cambridge, UK), rabbit antihuman AdipoR1 (5 g/ml; Bioss, London, UK), and rabbit antihuman AdipoR2 (5 g/ml; Bioss).

Techniques: In Vivo, Sampling, Injection, Immunohistochemistry, Staining