Journal: Molecular oncology

Article Title: The miR-200 family differentially regulates sensitivity to paclitaxel and carboplatin in human ovarian carcinoma OVCAR-3 and MES-OV cells.

doi: 10.1016/j.molonc.2015.04.015

Figure Lengend Snippet: Figure 1 e Characterization of OVCAR-3 GFP and OVCAR-3/TP GFP cell lines. (A) The cells were treated with different concentrations of paclitaxel (left panel) and carboplatin (right panel) during 72 h. Cells survival was measured by SRB assay. All data are expressed as the average percentage of survival values relative to an untreated control ± SD with significance determined between the indicated cell lines per paclitaxel or carboplatin concentration tested (*, P < 0.05; **, P < 0.01; ***, P < 0.001). (B) The constitutive expression of miR-200 family members was determined by real time PCR 48 h after cell seeding, using the RNU6 gene as an internal loading control, and calculating the ratio of parental to resistant cells. (C) The constitutive expression of CDH1, FN1 and VIM was measured in cells by using real time PCR 48 h after seeding. All data are expressed as the average of at least three measurements. Significance was determined between the OVCAR-3/TP GFP compared to the OVCAR-3 GFP cell line (*, P < 0.05; **, P < 0.01; ***, P < 0.001), (B and C). (D) EMT marker proteins were measured in cells 48 h after seeding using flow cytometry, and representative histograms of 10,000 events per cell line for each channel (E-cadherin-PE, Vimentin-Brilliant Violet 421, and Fibronectin-APC) are shown. Results for OVCAR-3 GFP cells are shown in orange and OVCAR-3/TP-GFP cells in blue.

Article Snippet: For intracellular staining, cells were exposed to Flow Cytometry Fixation Buffer (R & D Systems) for 20 min at 4 C in the dark, followed by the addition of Flow Cytometry Permeabilization buffer for 5 min at room temperature (R & D Systems).

Techniques: Sulforhodamine B Assay, Control, Concentration Assay, Expressing, Real-time Polymerase Chain Reaction, Marker, Cytometry