I am testing cancer prevention mechanisms and am going to dose HEK cells with ethidium bromide to cause cancer. What is the procedure for dosing with ethidium bromide?
Hello Emily Burdick
Ethidium bromide has been shown to bind strongly to both DNA and RNA in vivo and in vitro by intercalation between adjacent base pairs. Almost all studies to date implicate the mitochondrion as the primary target site of ethidium bromide at low concentrations. Nanogram to microgram concentrations of ethidium bromide per milliliter of culture medium induces loss of mitochondrial DNA and complete loss of mitochondrial DNA occurs after 5–6 weeks of exposure, and this approach has facilitated the study of mitochondrial mutations.
Treatment of cells with ethidium bromide has been found to result in the loss or alteration of mitochondrial DNA and RNA, the selective inhibition of mitochondrial DNA polymerase, the alteration of the cytochrome system, and morphological abnormalities of the mitochondrion. Studies in cells that have lost mitochondrial DNA over a period of weeks in culture report increased mRNA levels of genes involved in hypoxic response, mitochondrial ribosomal proteins, solute transport, and glycolytic pathways.
You may use concentrations ranging from 10 to 100 ng/ml for a few weeks.
You may want to refer to the articles attached below for more information.
https://www.frontiersin.org/journals/endocrinology/articles/10.3389/fendo.2014.00122/full
https://www.jbc.org/article/S0021-9258(19)42630-6/pdf
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