gw-5074 and Adenocarcinoma

Colorectal cancer (CRC) remains one of the leading causes of cancer-related death. Novel therapeutics are urgently needed, especially for tumours with activating mutations in. Colonosphere cultures were established from human tumour specimens obtained from patients who underwent colon or liver resection for primary or metastatic adenocarcinoma. The role of RAF1 was tested by generating knockdowns (KDs) using three independent shRNA constructs or by using RAF1-kinase inhibitor GW5074. Clone-initiating and tumour-initiating capacities were assessed by single-cell cloning and injecting CRC cells into immune-deficient mice. Expression of tight junction (TJ) proteins, localisation of polarity proteins and activation of MEK-ERK pathway was analysed by western blot, immunohistochemistry and immunofluorescence.. KD or pharmacological inhibition of RAF1 significantly decreased clone-forming and tumour-forming capacity of all CRC cultures tested, including. RAF1 plays a critical role in maintaining the transformed phenotype of CRC cells, including those with mutated

Topics: Adenocarcinoma; Animals; Cell Differentiation; Cell Polarity; Colorectal Neoplasms; Gene Expression; Gene Knockdown Techniques; Humans; Indoles; MAP Kinase Kinase Kinases; Mice; Neoplasm Transplantation; Phenols; Phosphorylation; Primary Cell Culture; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-raf; RNA, Small Interfering; Tight Junctions; Tumor Cells, Cultured