I am going over a reviewer's comments for my paper and being new to the research world I am not sure what the reviewer means when he/she is asking me whether the inhibitory effect of my product can be "rescued" for my cancer cell growth. He says I should do this to rule out toxicity. I did an MTT test, which did not show any effect of my product on the cell viability. Thanks in advance!
Hello Emilie,
According to the experiment, I believe the rescue is- if your drug is targetting an miRNA or protein target then you treat the cells with drug after inhibiting that protein/miRNA and then show whether it can rescue cells from drug effect. This will prove the specificity and possibility of that particular target in cells by drug.
Rest, MTT is also good and you can provide these two experiments for her reference..
Hope it helps !
Hi Emilie, your question is rahter general so be careful with the answers, there will be lots of assumptions. For me it sounds that the problem is that your product inhibits cell growth just as any other toxic compound would. MTT test was not convincing enough it seems. So you have to prove that the cells don`t just die due to the possible cytotoxic effects but the decline in growth rate is indeed due to the interference with the pathway/mechanism you hypothesize. So a resuce in this case I think would be an additional treatment that stimulate cell growth.
Ok my understanding of what you are saying is that the reviewer is asking for me to add something to the cells in order to encourage growth and see if my product still inhibits the growth? And I guess that something should be one of my targets?
Hello Emilie,
The question is very vague. It will be great if you can provide some details.
What was the experiment you performed when you talk about inhibitory effect..?
a little detail about the experiment will be greatly helpful in predicting what reviewer wants to know ..According to my experience, here rescue effect mean anything...like, if the inhibitory effect induced by drug can be reversed or rescued with the help of other compound....if the compound is toxic to normal cells (because if you can prove that, it will rule out its toxicity)..so u need to provide details dear ,,
Hope it helps !
I exposed breast cancer cells selected for a majority population of stem cells (also called mammospheres). I put my natural product in the medium (after filtering etc) and saw that with increasing dose fewer mammospheres grew (at a certain point almost none). I demonstrated that with MTT my product had no influence on the cell viability. Is this enough extra information? My product has a miRNA target and possible protein target.
Hello Emilie,
According to the experiment, I believe the rescue is- if your drug is targetting an miRNA or protein target then you treat the cells with drug after inhibiting that protein/miRNA and then show whether it can rescue cells from drug effect. This will prove the specificity and possibility of that particular target in cells by drug.
Rest, MTT is also good and you can provide these two experiments for her reference..
Hope it helps !
Hello Emile,
supposing you knock down (or better knockout)/inhibit your putative drug targets you should 1. See the same effect as with drug treatment and 2. Your drug shouldn't have additional effects. That wouldn't be really a rescue but would prove a point..
Another possibility would be overexpression of your targets to see if thereby you can decrease or even abrogate your drug's effects, i.e. rescue the cells.
To get into this further you could check for known markers (if there are any) of mamosphere formation and check if they decrease in the same manner with drug compared to knock down. Overexpression of these should then rescue your cells from the drug's effects.
However, as you want this paper out and if you addressed all other points sufficiently, the reviewer, as Ajay Kumar implied, might accept your MTT results as proof of lack of general toxicity.
Good Luck!
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