In order to analyse the effects of 5-fu on hct116 cells viability , I performed MTT assay(24h) for different doses of 5-fu(0.1mM to 80mM).
Based on the results of 24h MTT : viability of control were less than all doses .
Do you get the same results after repeating?
What concentration do you plate the cells at? The doubling time for hct119 is 21 hrs. In untreated sample, if the cell density is 0.5 million/well in a 6 well plate, then after 21 hrs, there are 1 million cells, and that is the highest capacity of each well in a 6 well plate. The cells will die in ctrl because now the nutrients also start running out in this well.. and the cells are now starved and hence cell death--> This can be one of the reasons you are getting higher cell death in control.
The time period of your experiment seems very very short. Also I have often found with plate assays that sometimes drug treatments for very short time periods often appear to increase apparent cell viability, this is often due to the mechanisms of the assay and disappear in longer assays.
Good Luck
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