Identification of EGFR and RAS Inhibitors using Caenorhabditis elegans
| Autor: | Thuy Nhu L Truong, Sabita Thapa, Ali Naji, Ransome van der Hoeven, Dharini van der Hoeven, John F. Hancock |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
General Immunology and Microbiology biology Effector General Chemical Engineering General Neuroscience Mutant Cancer medicine.disease biology.organism_classification Phenotype General Biochemistry Genetics and Molecular Biology Green fluorescent protein Cell biology 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Nematode medicine biology.protein Epidermal growth factor receptor 030217 neurology & neurosurgery Caenorhabditis elegans |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/61788 |
| Popis: | The changes in the plasma membrane localization of the epidermal growth factor receptor (EGFR) and its downstream effector RAS have been implicated in several diseases including cancer. The free-living nematode C. elegans possesses an evolutionary and functionally conserved EGFR-RAS-ERK MAP signal cascade which is central for the development of the vulva. Gain of function mutations in RAS homolog LET-60 and EGFR homolog LET-23 induce the generation of visible nonfunctional ectopic pseudovulva along the ventral body wall of these worms. Previously, the multivulval (Muv) phenotype in these worms has been shown to be inhibited by small chemical molecules. Here we describe a protocol for using the worm in a liquid-based assay to identify inhibitors that abolish the activities of EGFR and RAS proteins. Using this assay, we show R-fendiline, an indirect inhibitor of K-RAS, suppresses the Muv phenotype expressed in the let-60(n1046) and let-23(sa62) mutant worms. The assay is simple, inexpensive, is not time consuming to setup, and can be used as an initial platform for the discovery of anticancer therapeutics. |
| Databáze: | OpenAIRE |
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