Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Separation of cell survival, growth, migration, and mesenchymal transdifferentiation effects of fibroblast secretome on tumor cells of head and neck squamous cell carcinoma

doi: 10.1177/1010428317705507

Figure Lengend Snippet: FIB and SCC-25 cells produce TGF-β1 and the effects of TGF-β1 on SCC-25 cell proliferation and migration. (a) The TGF-β1 concentrations in the supernatants were measured using ELISA after 3 days of treatment. SCC-25 cells treated with 5 ng/mL TGF-β1 contained 917.1 ± 38.12 pg/mL in the supernatant. In the supernatant of the albumin-medium-treated (control) SCC-25 cells, there was almost no detectable TGF-β1 (0.3 ± 0.12 pg/mL). Significant levels of TGF-β1 were measured in the supernatants of FIB CM (164.1 ± 6.85 pg/mL) and mixed-culture CM (106.0 ± 5.82 pg/mL)-treated SCC-25 cells, whereas the TGF-β1 levels in co-culture (2.3 ± 0.94 pg/mL) were low. In the supernatant of SCC-25-CM-treated cells 10.4 ± 2.53 pg/mL TGF-β1 was measured. (b) After treatment with 5 ng/mL TGF-β1, there were significantly (p < 0.01) less SCC-25 cells growing than in the albumin-medium treated (control). (c) 0–96 h treatment with 5 ng/mL TGF-β1 leads to no change in coverage of the scraped area, while in the albumin-medium treated (control; p < 0.001), it was decreased. (d) 48–96 h treatment with 5 ng/mL TGF-β1 cells were also distributed over the scraped area (CM: conditioned medium; FIB: human gingival fibroblasts; **p < 0.01; ***p < 0.001). Cells treated with 5 ng/mL TGF-β1 showed elongated, mesenchymal-like morphology at 72–96 h (bars: 100 μm).

Article Snippet: Then, the cells were washed twice with DPBS and treated for 72 h with 2 mL/well albumin-supplied or CM by exchanges every 24 h. After completion of treatments, the supernatants were taken from the SCC-25 cells and frozen at −20°C to be used for the human TGF-β1 PicoKine™ enzyme-linked immunosorbent assay (ELISA Kit; Boster Biological Technology, Pleasanton, CA, USA) later on.

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Separation of cell survival, growth, migration, and mesenchymal transdifferentiation effects of fibroblast secretome on tumor cells of head and neck squamous cell carcinoma

doi: 10.1177/1010428317705507

Figure Lengend Snippet: Fold change EMT-related gene expressions in SCC-25 cells related to the albumin-medium-treated control.

Article Snippet: Then, the cells were washed twice with DPBS and treated for 72 h with 2 mL/well albumin-supplied or CM by exchanges every 24 h. After completion of treatments, the supernatants were taken from the SCC-25 cells and frozen at −20°C to be used for the human TGF-β1 PicoKine™ enzyme-linked immunosorbent assay (ELISA Kit; Boster Biological Technology, Pleasanton, CA, USA) later on.

Techniques:

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Separation of cell survival, growth, migration, and mesenchymal transdifferentiation effects of fibroblast secretome on tumor cells of head and neck squamous cell carcinoma

doi: 10.1177/1010428317705507

Figure Lengend Snippet: Fold change of vimentin and E-cadherin protein related to the control. The protein synthesis of vimentin, E-cadherin and β-actin in SCC-25 cells was determined using western blot analysis. β-actin was applied as a loading control. Fibroblasts (a, lane 7) served as positive control cells for vimentin, since this protein was contained in undetectable levels in parental SCC-25 cells. (a) Typical western blot of 57 kDa vimentin, 120 kDa E-cadherin, and 42 kDa β-actin in samples: (1) control SCC-25 treated with albumin medium; (2) SCC-25 treated with FIB CM; (3) SCC-25 treated with SCC-25 CM; (4) SCC-25 treated with mixed-culture CM; (5) SCC-25 co-cultured with fibroblasts; (6) SCC-25 treated with TGF-β1 (5 ng/mL); and (7) cultured fibroblasts, positive control for vimentin and negative control for E-cadherin (CM: conditioned medium). In FIB and mixed-culture CM-treated SCC-25 cells, a faint vimentin band appeared. (b–c) The band intensities were analyzed with densitometry. The different treatments were related to control cells treated with albumin-containing medium only. Densitometry graphs covered 10 comparable western blot experiments. (b) Two times upregulation of vimentin was shown by treatment with FIB CM. (c) E-cadherin showed decrease in co-culture and after TGF-β1 treatment.

Article Snippet: Then, the cells were washed twice with DPBS and treated for 72 h with 2 mL/well albumin-supplied or CM by exchanges every 24 h. After completion of treatments, the supernatants were taken from the SCC-25 cells and frozen at −20°C to be used for the human TGF-β1 PicoKine™ enzyme-linked immunosorbent assay (ELISA Kit; Boster Biological Technology, Pleasanton, CA, USA) later on.

Techniques: Western Blot, Positive Control, Cell Culture, Negative Control, Co-Culture Assay

Journal: Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine

Article Title: Separation of cell survival, growth, migration, and mesenchymal transdifferentiation effects of fibroblast secretome on tumor cells of head and neck squamous cell carcinoma

doi: 10.1177/1010428317705507

Figure Lengend Snippet: Comparison of the main changes observed in the secretome or co-culture treatments of tumor cells.

Article Snippet: Then, the cells were washed twice with DPBS and treated for 72 h with 2 mL/well albumin-supplied or CM by exchanges every 24 h. After completion of treatments, the supernatants were taken from the SCC-25 cells and frozen at −20°C to be used for the human TGF-β1 PicoKine™ enzyme-linked immunosorbent assay (ELISA Kit; Boster Biological Technology, Pleasanton, CA, USA) later on.

Techniques: Migration, Expressing