Journal: Oncology Letters

Article Title: Targeting CD146 in combination with vorinostat for the treatment of ovarian cancer cells

doi: 10.3892/ol.2017.5630

Figure Lengend Snippet: Induction of the adhesion molecule CD146 is a common phenomenon in vorinostat-treated ovarian cancer cells in vitro and in vivo . (A) A2780 and SKOV3 cells were treated with 2.5 µmol/l vorinostat or 500 nmol/l TSA for 12 h and subjected to analysis of quantitative polymerase chain reaction for the mRNA levels of CD146. Results are normalized to those of 18s RNA and expressed as the fold induction compared with the DMSO-treated group (*P<0.05). (B) A2780 and SKOV3 cells were treated with 2.5 µmol/l vorinostat for 24 h and analyzed for the protein levels of CD146 by western blot analysis. (C) A2780 cells were treated with 2.5 µmol/l vorinostat or DMSO for 12 h and were then analyzed by immunofluorescent analysis for staining of CD146, obtaining representative images under a confocal microscope (magnification, ×600). (D) SKOV3 tumor-bearing mice were treated with 25 mg/kg of vorinostat or DMSO for 24 h and CD146 expression was determined by immunohistochemistry and quantified by laser scanning cytometry. Images represent typical data (Total positive rate for CD146 (SP plus P) is the mean ± standard deviation (n=10). CD146, cluster of differentiation 146; DMSO, dimethyl sulfoxide; TSA, trichostatin A; SP, strong positive; P, positive; N, negative; IHC, immunohistochemistry; mRNA, messenger RNA; LSC, laser scanning cytometry.

Article Snippet: Fluorescein isothiocyanate-conjugated mouse anti-human CD146 mAb (1:100; 11-1469-42) was purchased from eBioscience, Inc. San Diego, CA, USA).

Techniques: In Vitro, In Vivo, Real-time Polymerase Chain Reaction, Western Blot, Staining, Microscopy, Expressing, Immunohistochemistry, Cytometry, Standard Deviation

Journal: Oncology Letters

Article Title: Targeting CD146 in combination with vorinostat for the treatment of ovarian cancer cells

doi: 10.3892/ol.2017.5630

Figure Lengend Snippet: Effect of vorinostat on the expression of CD146 in clinical tumor samples.

Article Snippet: Fluorescein isothiocyanate-conjugated mouse anti-human CD146 mAb (1:100; 11-1469-42) was purchased from eBioscience, Inc. San Diego, CA, USA).

Techniques: Expressing, Biomarker Discovery

Journal: Oncology Letters

Article Title: Targeting CD146 in combination with vorinostat for the treatment of ovarian cancer cells

doi: 10.3892/ol.2017.5630

Figure Lengend Snippet: Targeting CD146 substantially enhanced vorinostat-induced killing in ovarian cancer cells. (A) Ovarian cancer A2780 cells were cultured with control mAb mIgG or AA98, which has been confirmed to significantly knockdown the expression of CD146 by western blot analysis. (B) A2780/SKOV3/Caov3 ovarian cancer cells were exposed to 2.5 µmol/l vorinostat for 72 h and subjected to apoptosis assay for the determination of their drug sensitivity, where each data point represents the mean ± standard error of the mean of 3 replicates. (C) A2780 or (D) SKOV3 cells were treated with 10 µg/ml mAb AA98 or 2.5 µmol/l vorinostat + isotype-matched mIgG, or vorinostat + mAb AA98 for 24 h and then subjected to the soft agar colony-forming assay (*P<0.05). Results are expressed as a percentage of colonies in the DMSO-treated group. Each data point represents the mean ± standard deviation of 3 replicates. CD146, cluster of differentiation 146; mAb, monoclonal antibody; mIgG, monoclonal immunoglobulin G; DMSO, dimethyl sulfoxide; SAHA, suberoylanilide hydroxamic acid.

Article Snippet: Fluorescein isothiocyanate-conjugated mouse anti-human CD146 mAb (1:100; 11-1469-42) was purchased from eBioscience, Inc. San Diego, CA, USA).

Techniques: Cell Culture, Control, Knockdown, Expressing, Western Blot, Apoptosis Assay, Standard Deviation

Journal: Oncology Letters

Article Title: Targeting CD146 in combination with vorinostat for the treatment of ovarian cancer cells

doi: 10.3892/ol.2017.5630

Figure Lengend Snippet: Knockdown of CD146 promotes vorinostat-induced apoptosis via suppression of the Akt pathway in ovarian cancer cells. (A) A2780 cells were treated as depicted for 24 h (5 µmol/l vorinostat; 10 µg/ml mAb AA98) and examined for protein levels of total Akt, p-Akt, P-4E-BP1, p-S6K1, p-GSK-3β and β-actin. VT analysis by western blotting. (B) A2780 cells stably transfected with the AAkt2 plasmid were treated as depicted for 24 h and examined for protein levels of total Akt and p-Akt by western blot analysis. A2780 cells stably transfected with pcDNA3.1 plasmid were treated as control group. (C) A2780 cells were treated with as depicted for 24 h (5 µmol/l vorinostat; 10 µg/ml mAb AA98; 5 µmol/l triciribine) and examined for protein levels of total Akt and p-Akt by western blot analysis. CD146, cluster of differentiation 146; Akt, protein kinase B; p-, phosphorylated; 4E-BP1, 4E-binding protein 1; S6K1, ribosomal protein S6 kinase-1; GSK-3β, glycogen synthase kinase 3β; VT, vorinostat treatment; DMSO, dimethyl sulfoxide.

Article Snippet: Fluorescein isothiocyanate-conjugated mouse anti-human CD146 mAb (1:100; 11-1469-42) was purchased from eBioscience, Inc. San Diego, CA, USA).

Techniques: Knockdown, Western Blot, Stable Transfection, Transfection, Plasmid Preparation, Control, Binding Assay

Journal: Oncology Letters

Article Title: Targeting CD146 in combination with vorinostat for the treatment of ovarian cancer cells

doi: 10.3892/ol.2017.5630

Figure Lengend Snippet: Targeting CD146 synergized with vorinostat to substantially inhibit ovarian cancer growth. (A) When the tumor reached a diameter of 5–6 mm, SKOV3 tumor-bearing mice were grouped (n=10) and administered intraperitoneally with PBS (black), 8 mg/kg mAb AA98 (green), 20 mg/kg vorinostat plus mIgG (brown), or vorinostat plus mAb AA98 (blue). Mean tumor volumes were monitored at specific time points subsequent to treatment. Each data point represents mean ± standard deviation (*P<0.05; n=10). (B) Kaplan-Meier survival curves of SKOV3 tumor-bearing mice following various treatments as indicated. Each group consisted of 10 animals. PBS served as a control. CD146, cluster of differentiation 146; mAb, monoclonal antibody; mIgG, monoclonal immunoglobulin G; SAHA, suberoylanilide hydroxamic acid.

Article Snippet: Fluorescein isothiocyanate-conjugated mouse anti-human CD146 mAb (1:100; 11-1469-42) was purchased from eBioscience, Inc. San Diego, CA, USA).

Techniques: Standard Deviation, Control