Journal: Oncology Reports
Article Title: Synergistic activity of agents targeting growth factor receptors, CDKs and downstream signaling molecules in a panel of pancreatic cancer cell lines and the identification of antagonistic combinations: Implications for future clinical trials in pancreatic cancer
doi: 10.3892/or.2020.7822
Figure Lengend Snippet: Effect of afatinib, dinaciclib, dasatinib, stattic and NVP-AEW742 with or without ligands (EGF, HB-EGF, IGF-II) on the phosphorylation of EGFR and downstream cell signaling molecules including MAPK, AKT, STAT3, SRC and IGF-IR in BxPC-3 (A) and Capan-1 (B) cells. The cells were cultured in 10% FBS RPMI-1640 medium to near confluency. Cells were washed once with 0.5% FBS RPMI-1640 medium and incubated with selected agents (400 nM) for 1 h and then stimulated with 40 nM ligands (EGF, HB-EGF and IGF-II) for 15 min. Cells were then lysed, separated using SDS-PAGE, transferred onto PDVF membranes, probed with the antibodies of interest and visualized using LI-COR software. EGF, epidermal growth factor; HB-EGF, heparin-binding EGF-like growth factor; IGF-II, insulin-like growth factor II; EGFR, epidermal growth factor receptor; MAPK, mitogen-activated protein kinase; AKT, protein kinase B or PKB; STAT3, signal transducer and activator of transcription 3; SRC, proto-oncogene tyrosine kinase SRC; IGF-IR, insulin-like growth fact) or 1 receptor.
Article Snippet: The antibodies for flow cytometry including mouse anti-EGFR (HM43.16B) and anti-HER2 (HM50.67A) were raised in-house against the external domain of these receptors ( ) whereas mouse anti-HER3 (MAB3481), anti-HER4 (MAB11311), ALK7 (MAB77491), HGF R/c-MET (MAB3582), PDGFRα (MAB1264), PDGFRβ (MAB1263) and IGF-IR (MAB391) were purchased from R&D Systems (Europe Ltd. UK) and Insight Biotechnology (Middlesex, UK), respectively.
Techniques: Phospho-proteomics, Cell Culture, Incubation, SDS Page, Software, Binding Assay
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