human bc cell lines t24  (ATCC)

Journal: Medical Sciences

Article Title: Leptin Drives Breast Cancer Aggressiveness Acting Through the Activation of the NCOA1/STAT3 Pathway

doi: 10.3390/medsci14010032

Figure Lengend Snippet: A high leptin dose, mimicking hyperleptinemia in individuals with obesity, is exclusively able to enhance BC cells proliferation. ( a ) LEPR expression levels in MCF-7 and MDA-MB-231 cells were quantified by flow cytometry, and the effect of leptin (10 and 100 ng/mL) on LEPR expression in both cell lines was assessed after 48 h. Mean fluorescence intensity (MFI) was quantified from flow cytometry data. ( b ) MCF-7 and MDA-MB-231 proliferation was assessed after 48 h treatment with 10 or 100 ng/mL of leptin using an MTT assay. ( c ) Colony formation ability of MCF-7 and MDA-MB-231 cells was evaluated after 14 days of culture. Colony area was quantified, and data are presented as fold change relative to the control. ( d ) Cyclin D1 expression levels in MCF-7 and MDA-MB-231 cells were determined by qRT-PCR after treatment with 10 or 100 ng/mL of leptin for 48 h. Data were normalized to endogenous GAPDH mRNA levels. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The human BC cell lines MCF-7 (RRID:CVCL_0031) and MDA-MB-231 (RRID:CVCL_0062) were obtained from American Type Cell Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Flow Cytometry, Fluorescence, MTT Assay, Control, Quantitative RT-PCR

Journal: Medical Sciences

Article Title: Leptin Drives Breast Cancer Aggressiveness Acting Through the Activation of the NCOA1/STAT3 Pathway

doi: 10.3390/medsci14010032

Figure Lengend Snippet: Elevated leptin levels promote BC cell 2D and 3D migration. MCF-7 and MDA-MB-231 cells were treated with 10 or 100 ng/mL of leptin and subjected to ( a ) wound-healing and ( b ) spheroid dissemination assays. Images were captured at 0 h and 48 h post-treatment. Cell migration was quantified. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The human BC cell lines MCF-7 (RRID:CVCL_0031) and MDA-MB-231 (RRID:CVCL_0062) were obtained from American Type Cell Collection (ATCC, Manassas, VA, USA).

Techniques: Migration

Journal: Medical Sciences

Article Title: Leptin Drives Breast Cancer Aggressiveness Acting Through the Activation of the NCOA1/STAT3 Pathway

doi: 10.3390/medsci14010032

Figure Lengend Snippet: High leptin levels stimulate EMT in BC cells. ( a ) Representative phase-contrast microscopy images illustrating morphological changes in MCF-7 and MDA-MB-231 cells following 48 h treatment with 100 ng/mL of leptin. ( b ) Representative confocal microscopy images of F-Actin immunofluorescence in MCF-7 cells treated with 100 ng/mL leptin for 48 h. ( c ) Representative confocal microscopy images of E-Cadherin immunofluorescence in MCF-7 cells treated with 100 ng/mL leptin for 48 h. Scale bar: 20 μm. Scatter plots showing correlation of EMT SCORE with ( d ) LEP and ( e ) LEPR expression in The Breast Invasive Carcinoma (TCGA, PanCancer Atlas) dataset.

Article Snippet: The human BC cell lines MCF-7 (RRID:CVCL_0031) and MDA-MB-231 (RRID:CVCL_0062) were obtained from American Type Cell Collection (ATCC, Manassas, VA, USA).

Techniques: Microscopy, Confocal Microscopy, Immunofluorescence, Expressing

Journal: Medical Sciences

Article Title: Leptin Drives Breast Cancer Aggressiveness Acting Through the Activation of the NCOA1/STAT3 Pathway

doi: 10.3390/medsci14010032

Figure Lengend Snippet: Elevated leptin dose triggers increased STAT3 expression and phosphorylation. ( a ) STAT3 gene expression levels in MCF-7 and MDA-MB-231 cells were assessed by qRT-PCR following 24 h treatment with 10 or 100 ng/mL of leptin. ( b ) Representative Western blot showing total STAT3 and phosphorylated STAT3 protein levels in MCF-7 and MDA-MB-231 cells treated with different leptin concentrations (10 and 100 ng/mL) for 24 h. Band intensities were quantified using ImageJ software and normalized to β-actin levels. ( c ) MCF-7 were subjected to wound healing assay following treatment with 100 ng/mL of leptin, with or without 100 µM of AG490 (STAT3 inhibitor). Images were captured at 0 h and 48 h post-treatment. Cell migration was quantified, and the recovery percentage was calculated. ( d ) Representative confocal microscopy images of E-Cadherin immunofluorescence in MCF-7 cells treated with 100 ng/mL of leptin, in the presence or absence of AG490 (100 µM). Scale bar: 20 μm. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The human BC cell lines MCF-7 (RRID:CVCL_0031) and MDA-MB-231 (RRID:CVCL_0062) were obtained from American Type Cell Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Phospho-proteomics, Gene Expression, Quantitative RT-PCR, Western Blot, Software, Wound Healing Assay, Migration, Confocal Microscopy, Immunofluorescence

Journal: Medical Sciences

Article Title: Leptin Drives Breast Cancer Aggressiveness Acting Through the Activation of the NCOA1/STAT3 Pathway

doi: 10.3390/medsci14010032

Figure Lengend Snippet: Increased expression of nuclear coactivator 1 ( NCOA1 ) was induced by elevated leptin doses and was found to be positively correlated with LEP , LEPR , and STAT3 target gene expression in BC patients. ( a ) ERα , ERRα , ERRγ , and NCOA1 gene expression levels in MCF-7 and MDA-MB-231 cells were assessed by qRT-PCR following treatment with 100 ng/mL of leptin for 24 h. Data were normalized to endogenous GAPDH mRNA levels. ( b ) MCF-7 cells were transduced with Ad-NCOA1, and 48 h later, mRNA levels of 2 STAT3 target genes, VEGF and Cyclin D1 , were measured by qRT-PCR. * p < 0.05; ** p < 0.01; *** p < 0.001. ( c ) Protein–protein interaction (PPI) network of LEP, LEPR, NCOA1, STAT3, and related genes. GeneMANIA plot revealing that these genes were linked by physical interaction at 77.64%. Scatter plots showing correlation of NCOA1 with ( d ) LEP and ( e ) LEPR expression in The Breast Invasive Carcinoma TCGA PanCancer Atlas dataset. Correlation of NCOA1 with ( f ) STAT3-gene signature and ( g ) P-STAT3-gene signature in the same dataset. Pearson’s correlation coefficient was applied to assess the association between variables, with p values < 0.05 considered statistically significant.

Article Snippet: The human BC cell lines MCF-7 (RRID:CVCL_0031) and MDA-MB-231 (RRID:CVCL_0062) were obtained from American Type Cell Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Targeted Gene Expression, Gene Expression, Quantitative RT-PCR, Transduction