Hello;
I've done MTT assays to investigate whether my peptide drug could inhibit tumor cell proliferation. I used MDA-MB-231 as positive cells and MCF-7 as negative ones which do not express target protein. It could be interpreted from MTT assay results that my peptide could decrease cell viability in positive cells but did not effect negative control cells in the same way, in other word no decrease in viability was observed in MCF-7 cells in none of the tested concentrations. So I concluded that my peptide shows anti-proliferative properties.
However, I doubt whether viability reduction was due to really manipulating downstream proliferation pathways or it was only because of peptide direct cytotoxicity against MDA-MB-231 cells.
Is there anyway to distinguish between these two events?
Thanks in advance!
I had the same question. A good discussion on RG here:
https://www.researchgate.net/post/How_to_distinguish_by_any_assay_between_antiproliferative_and_cytotoxiicty_of_cell_line_mda-mb231_caused_by_chemical
Dear Rajive, you should use MCF10A ( a normal breast cancer cell) cells along with MDA-MB-231 for MTT assay with your test agent. This will help to get the specific effect on cancer cells.
Dear,Raheleh
MTT assay can indicate total anti-growth effects of a drug which includes both cytotoxic ant anti-proliferative effects. Thus, to discriminate you can first determine the cytotoxic activity of your drug at a given concentration using LDH assay or even the more available and convenient trypan blue test. Then, run an MTT assay using the same concentrations and incubation times. After obtainung these two data for each concentration, you can discriminate by subtracting cytotoxic value from total growth-inhibition value to obtain anti-proliferative effect at that dose. I used the same protocol in our paper. You can read the paper to get more information on the issue. Here is the link: https://pubmed.ncbi.nlm.nih.gov/29266560/
Hope this can help you.
Good luck
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