Journal: Breast Cancer : Targets and Therapy

Article Title: Expression and functionality of TRPV1 in breast cancer cells

doi: 10.2147/BCTT.S121610

Figure Lengend Snippet: Expression profiles of TRP channels in the SUM149PT breast cancer cell line. Notes: ( A ) NGS-based RNA-Seq analysis of the expression of TRP channels. ( B ) Validation of the expression of TRPV1 in SUM149PT cells via RT-PCR. +, +RT, cDNA; −, −RT, RNA; g, genomic DNA as a control. ( C ) Immunocytochemical staining of SUM149PT cells with specific TRPV1 antibodies. Left: SUM149PT cells. Right: Negative control: SUM149PT cells stained with secondary antibody alone. Staining of the cell nuclei was performed using DAPI (blue). Scale bar: 10 µm. Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; M, marker; NGS, next-generation sequencing; PCR, polymerase chain reaction; RT, reverse transcriptase; TRP, transient receptor potential.

Article Snippet: SUM149PT cells were incubated at 37°C for 30 min with Ringer’s solution containing 3 mM FURA-2-AM (fura-2-acetoxymethyl ester; Molecular Probes, Eugene, OR, USA).

Techniques: Expressing, RNA Sequencing, Biomarker Discovery, Reverse Transcription Polymerase Chain Reaction, Control, Staining, Negative Control, Marker, Next-Generation Sequencing, Polymerase Chain Reaction, Reverse Transcription

Journal: Breast Cancer : Targets and Therapy

Article Title: Expression and functionality of TRPV1 in breast cancer cells

doi: 10.2147/BCTT.S121610

Figure Lengend Snippet: Functional characterization of TRPV1 activation by capsaicin in SUM149PT cells. Notes: ( A ) Representative image of SUM149PT cells stimulated with capsaicin (10 µM) in calcium imaging analysis. To ensure the viability of the cells, ATP (10 µM) was applied finally and served as a positive control. N > 3 with n = 12 measurements in six cell culture dishes with ~200 cells. ( B ) Activation of SUM149PT cells upon repetitive capsaicin application (10 µM). ( C ) Analysis of cell proliferation via CyQuant proliferation assay after capsaicin stimulation for 48 h (150 µM); n = 20. ( D ) Analyses of cell migration by scratch assay after capsaicin (150 µM) stimulation for 48 h. Bar chart showing statistical analysis of the area overgrown in scratch assay experiments; n = 3 assays. ( E ) Annexin V-FITC staining of SUM149PT cells stimulated by capsaicin (10 and 150 µM). Early apoptotic cells (Annexin V positive, PI negative) are shown in green; necrotic cells (Annexin V positive, PI positive) are shown in yellowish-red, and viable cells are shown in bright phase (Annexin V negative, PI negative). Bar chart showing statistical analysis of cells undergoing necrosis; n = 3 stains of five slices with ~300 cells. ( F ) Calcium signal upon application of capsaicin (100 µM) and capsaicin (100 µM) + capsazepin (50 µM); n = 6 cell culture dishes with ~200 cells. The shaded area represents the application of the antagonist capsazepin. ( G ) Analysis of the cell proliferation via CyQuant proliferation assay after capsaicin (50 µM) and capsaicin (50 µM) + capsazepin (50 µM) stimulation for 48 h; n = 10. * P <0.05, *** P <0.001. Scale bar = 10 µm. Abbreviations: C, capsaicin; CZ, capsazepin; FITC, fluorescein isothiocyanate; PI, propidium iodide; F340/F380, ratio of the fluorescence intensities at 340 nm and 380 nm excitation; C, capsaicin; CZ, capsazepin; ATP, adenosine triphosphate; ns, not significant.

Article Snippet: SUM149PT cells were incubated at 37°C for 30 min with Ringer’s solution containing 3 mM FURA-2-AM (fura-2-acetoxymethyl ester; Molecular Probes, Eugene, OR, USA).

Techniques: Functional Assay, Activation Assay, Imaging, Positive Control, Cell Culture, CyQUANT Assay, Proliferation Assay, Migration, Wound Healing Assay, Staining, Fluorescence