Demonstration by real-time polymerase chain reaction that cellular DNA alkylation by novel aminoindoline compounds affects expression of the protooncogene c-myc
| Autor: | Lynnette R. Ferguson, William A. Denny, Stephanie M. Nelson |
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| Rok vydání: | 2005 |
| Předmět: |
Indoles
Time Factors Alkylation Molecular Conformation Biology Toxicology law.invention Cell Line Proto-Oncogene Proteins c-myc chemistry.chemical_compound Duocarmycins Downregulation and upregulation law Cell Line Tumor medicine Humans Urea Polymerase chain reaction Reverse Transcriptase Polymerase Chain Reaction General Medicine DNA U937 Cells Molecular biology In vitro Reverse transcriptase DNA Alkylation Real-time polymerase chain reaction Mechanism of action chemistry RNA medicine.symptom |
| Zdroj: | Chemical research in toxicology. 18(2) |
| ISSN: | 0893-228X |
| Popis: | Aminoindolines, analogues of the potent DNA alkylating agent seco-CBI-TMI, bind to and alkylate in the minor groove of AT-rich DNA in vitro. Here we extend the in vitro mechanism of action studies by treating cells in culture and examining the DNA binding patterns within AT-rich regions of the protooncogene locus c-myc, using a real-time polymerase chain reaction (PCR) stop assay. In addition, real-time reverse transcriptase (RT) PCR is used to examine the immediate effects of drug treatment on c-myc expression. These analyses demonstrate a concentration and time dependence for DNA alkylation at the chosen sites within the c-myc locus, as well as a prompt and significant downregulation of c-myc expression. While downregulation of this important growth regulator is likely not the only consequence of aminoindoline treatment, these studies begin to address the cellular pathways that are involved in the potent cytotoxic effects observed and provide insights for the future development of anticancer drugs of this class. |
| Databáze: | OpenAIRE |
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