Publicação
Characterization of in vitro models for the study of candidate G-quadruplex ligands targeting the human c-KIT proto-oncogene promoter
| Resumo: | Proto-oncogene c-KIT has been implicated in the development and growth of several tumors, e.g. mast cell tumors (MCT) in dogs and gastrointestinal stromal tumors (GIST) in humans. Several therapeutic approaches directed to the blocking of receptor tyrosine kinases (RTK), such as c-KIT, have been created. However, after a short period of recovery, these drugs lose efficiency and the tumor relapses. A new approach, aiming to control c-KIT’s transcription, is being tested. This approach relies on the use of small molecule inhibitors (SMI) that specifically block DNA secondary structures, G-quadruplexes, located on the promoter regions of many proto-oncogenes, including c-KIT. The main goal of this work is the development of in vitro models through which the study of candidate SMIs for human c-KIT is possible. An in vitro model, composed by cytotoxicity tests aimed for the determination of the SMI’s inhibitory concentration 50 (IC50) on two human cell lines and by real time quantitative PCR (qPCR) for the study of gene expression alterations, has been developed and validated. The cytotoxicity tests were also used to identify the IC50 of three candidate ligands for c-KIT. |
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| Autores principais: | Shahidian, Lara Zorro |
| Assunto: | c-KIT cytotoxicity G-quadruplex HGC27 MCF7 SMI citotoxicidade pequenas moléculas inibidoras |
| Ano: | 2013 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Proto-oncogene c-KIT has been implicated in the development and growth of several tumors, e.g. mast cell tumors (MCT) in dogs and gastrointestinal stromal tumors (GIST) in humans. Several therapeutic approaches directed to the blocking of receptor tyrosine kinases (RTK), such as c-KIT, have been created. However, after a short period of recovery, these drugs lose efficiency and the tumor relapses. A new approach, aiming to control c-KIT’s transcription, is being tested. This approach relies on the use of small molecule inhibitors (SMI) that specifically block DNA secondary structures, G-quadruplexes, located on the promoter regions of many proto-oncogenes, including c-KIT. The main goal of this work is the development of in vitro models through which the study of candidate SMIs for human c-KIT is possible. An in vitro model, composed by cytotoxicity tests aimed for the determination of the SMI’s inhibitory concentration 50 (IC50) on two human cell lines and by real time quantitative PCR (qPCR) for the study of gene expression alterations, has been developed and validated. The cytotoxicity tests were also used to identify the IC50 of three candidate ligands for c-KIT. |
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