Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Resistance to multikinase inhibitor actions mediated by insulin like growth factor-1

doi: 10.1186/s13046-015-0210-1

Figure Lengend Snippet: Antagonism by IGF1 of Regorafenib-mediated growth inhibition of HCC cell Lines. a . Hep3B, PLC/PRF/5 and HepG2 cells were cultured in presence of Regorafenib 1–5 μM, IGF1 40 ng/ml, GSK1838705A 1 μM. MTT assay was assessed after 48 h (48 h). b . PLC/PRF/5 cells were cultured in presence of Regorafenib 5 μM and IGF1 40 ng/ml in different time conditions. In the first group (light gray) the cells previously treated with IGF1 for 48 h received Regorafenib for the following 24 h. In the second group (dark gray) the cells, previously treated with Regorafenib for 24 h, received IGF for the following 48 h. MTT assay was assessed after 72 h. c . PLC/PRF/5 cells were synchronized in the S phase of the cell cycle using thymidine 0.2 M (T0), after 6 h from block release (T1), the cells were processed with the Cell Cycle Kit and analyzed with Muse Cell Analyzer to evaluate the percentage of cells in G0/G1, S and G2/M phases. The panels represent an example of DNA content profile in different treatment conditions. The mean of three independent experiments was plotted in the relative graph, where the values are calculated as fold increase of the cells in G2/M at T1 respect to control cells at T0. The results of three independent experiments are expressed as means ± SD. *** p < 0.0001

Article Snippet: The Human IGF1 ELISA kit (Wuhan Boster Biological Technology LTD, Wuhan, China) was used for the in vitro quantitative determination of human IGF1 in FBS (control) and serial dilution of hPL, according to the user’s guide.

Techniques: Inhibition, Cell Culture, MTT Assay, Blocking Assay, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Resistance to multikinase inhibitor actions mediated by insulin like growth factor-1

doi: 10.1186/s13046-015-0210-1

Figure Lengend Snippet: Antagonism by IGF1 of Regorafenib-mediated induction of apoptosis. a . PLC/PRF/5 cell line cultured in 1 % FBS medium was treated with IGF1 40 ng/ml in combination with Regorafenib 5 μM and GSK 1 μM. The Muse Annexin V kit was used to evaluate the percentage of apoptotic cells. The means ± SD of three independent experiments is plotted in the relative graph. *** p < 0.0001. b . Representative Western blot that shows anti-apoptotic (phospho-survivin, Bcl-xL and Bcl-2) and pro-apoptotic (Bim, truncated-Bid and Bad) proteins in PLC/PRF/5 cells treated as described above

Article Snippet: The Human IGF1 ELISA kit (Wuhan Boster Biological Technology LTD, Wuhan, China) was used for the in vitro quantitative determination of human IGF1 in FBS (control) and serial dilution of hPL, according to the user’s guide.

Techniques: Cell Culture, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Resistance to multikinase inhibitor actions mediated by insulin like growth factor-1

doi: 10.1186/s13046-015-0210-1

Figure Lengend Snippet: Antagonism by IGF1 of regorafenib-mediated inhibition of migration and invasion. PLC/PRF/5 cell line cultured in 1 % FBS medium was treated with IGF1 40 ng/ml alone or in combination with Regorafenib 5 μM and GSK 1 μM. a . Migration assay was performed as described and the microscopic analysis was assessed at the time of the scratch (T0) and after 48 h (T2). The values were expressed as percentage of migration, where 100 % represents the scratch completely closed. b . The percentage of invasion was calculated comparing the invading drug-treated cells to drug-untreated control cells (100 %). The results of three independent experiments are expressed as means ± SD. *** p < 0.0001

Article Snippet: The Human IGF1 ELISA kit (Wuhan Boster Biological Technology LTD, Wuhan, China) was used for the in vitro quantitative determination of human IGF1 in FBS (control) and serial dilution of hPL, according to the user’s guide.

Techniques: Inhibition, Migration, Cell Culture, Control

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Resistance to multikinase inhibitor actions mediated by insulin like growth factor-1

doi: 10.1186/s13046-015-0210-1

Figure Lengend Snippet: Effects of IGF1 receptor (IGFR1) inhibitor (GSK1838705A) on Regorafenib-mediated growth inhibition in PLC/PRF/5 cells. PLC/PRF/5 cells were cultured in 1 % FBS medium in presence of IGF1 40 ng/ml or platelet lysate corresponding to 3.75 × 10 7 platelets/ml, Regorafenib 5 μM and GSK1838705A 1 μM, using the conditions indicated in the graph. MTT assay was assessed after 48 h. The results of three independent experiments are expressed as mean ± SD. ** p < 0.001; *** p < 0.0001

Article Snippet: The Human IGF1 ELISA kit (Wuhan Boster Biological Technology LTD, Wuhan, China) was used for the in vitro quantitative determination of human IGF1 in FBS (control) and serial dilution of hPL, according to the user’s guide.

Techniques: Inhibition, Cell Culture, MTT Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Resistance to multikinase inhibitor actions mediated by insulin like growth factor-1

doi: 10.1186/s13046-015-0210-1

Figure Lengend Snippet: MAPK signaling and IGFR1 expression in PLC/PRF/5 cells. a . Western blot of MAPK proteins that shows IGF1 antagonism effect on Regorafenib action in PLC/PRF/5 cells. b . Activated IGF1 receptor (IGFR1) after treatment with Regorafenib (R), IGF1, IGF1 + R, IGF1 + GSK and IGF1 + R + GSK, respectively. c . IGF1 effect on the BRAF protein that is direct Regorafenib target

Article Snippet: The Human IGF1 ELISA kit (Wuhan Boster Biological Technology LTD, Wuhan, China) was used for the in vitro quantitative determination of human IGF1 in FBS (control) and serial dilution of hPL, according to the user’s guide.

Techniques: Expressing, Western Blot