Molecular Characterisation of Ret Tyrosine Kinase Signalling
نویسندگان
چکیده
....................................................................... 10 1. REVIEW OF THE LITERATURE ........................................ 11 1.1 RET RECEPTOR TYROSINE KINASE.................................. 11 1.1.1 RET gene and RET proteins............................................. 11 1.1.2 RET is a functional receptor for GDNF-family ligands (GFLs).... 12 1.1.3 RET-dependent GFLs signaling........................................ 14 1.1.4 RET-independent GDNF/GFRα1 signaling.......................... 17 1.1.5 Activation of RET by other growth factors............................ 18 1.1.6 Role of RET signaling during development.......................... 19 1.2 RET INTERACTINGPROTEINS.........................................20 1.2.1 Downstream signalling pathways...................................... 22 1.2.2 Shc family adaptor proteins.............................................. 23 1.3 RET ACTIVATION IN HUMAN CANCER............................. 25 1.3.1 RET in papilary carcinomas of the thyroid gland.................... 25 1.3.2 Other genes activated in papilary thyroid carcinomas (PTCs)..... 30 1.3.3 Medulary thyroid carcinoma............................................ 30 1.3.4 RET in MEN 2 syndromes............................................... 31 1.3.5 RET in sporadic medulary thyroid carcinoma (sMTC)............. 36 1.3.6 Diagnosis and management of MEN 2................................. 38 1.4 RET GERMLINE INACTIVATING MUTATIONS IN HIRSCHSPRUNG’S DISEASE................................................. 40 2. AIMS OF THE PRESENT STUDY......................................... 45 3. MATERIALS AND METHODS............................................. 46 3.1 Cel lines ....................................................................... 46 3.2 Antibodies,growth factors,recombinant proteins,chemical inhibitors.. 46 3.3 DNA constructs .............................................................. 47 3.4 Transfection of cel lines ................................................... 48 3.5 Adenovirus construction .................................................... 49 3.6 Immunoprecipitations ........................................................ 49 3.7 GST proteins .................................................................. 50 3.8 Western bloting.............................................................. 50 3.9 In vitro kinase assays ........................................................ 50 3.10 Biotinylation of cel surface proteins .................................... 50 3.11 I-labeled GDNF and HGF binding, chemical cross-linking........ 50 3.12 Cel migration and chemotaxis assays ................................... 51 3.13 Branching tubule formation assay in colagen gel ..................... 51 3.14 Cel apoptosis assays ....................................................... 51 3.15 Cel proliferation assays................................................... 51 3.16 Anchorage-independent growth assay ................................... 51 3.17 Kidney cultures .............................................................. 51
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