Hi researchers, I was searching for the synergism of two different drugs by running mtt assay,I analyzed my data and I found them synergism, so I used each drug IC50 and both Ic50 to check them by Anexin V/PI kit, but in flowcytometry I found necrosis more than apoptosis and also I found more apoptosis in each one treated with one drug instead of two drug, How is it possible? Why does necrosis happen? and why aren't they synergism in flowcytometry?
I used flowcytometry to run each IC50 for the different time of 24, 48 and 72 hours.
1. It could be secondary necrosis: an early time point such as 8 hrs post treatment could help to see apoptosis in combination condition.
2. Could be real necrosis due to hyper ROS production or due to p53 impairment.
3. Try caspase-3 inhibitor with combination condition; which could reduce necrosis.
4. may be the cells express more MLKL in combination treatment so that it shifts the balance towards necroptosis.
probably you need to decrease the dose in combination.
it seems the ic50 of combination is not the same as their ic50 when you use it
individually.
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