Hi,
i am working on effects of gemcitabine in HepG2 cell line and i should use the IC50 of Gemcitabine in the experiments.i found different values in literature and the numbers are highly variable ranging from lower nanomolar to lower micromolar for the same cell line under same conditions.!!!!!!
but in my tests,which i use different dose of gemcitabine from 0.25 to 16 micro molar ,from dose up to 1 micro molar ,i got same viability percentage.(60 percent)for all of drug doses.and also when i look at my cells,their morphology change a little bit and cells become wider like they are in G1 phase of cell cycle and the apoptotic cells are not exist in my plates.i think that the drug arrest my cells proliferation.i cant get the IC50.
is it normal ???
what should i do?
Dear Shadi Zerehpoosh
Please refer to this website https://www.cancerrxgene.org to find IC50 of drugs in cancer cell lines. first, try to expand your rang of treatments for example from 100 micro molar to 1 nano molar because you will be able to obtain better results using GraphPad Prism software. Second, it is better to use 5 or 10 fold changes between your serial dilutions.
third, some of drugs react with MTT and increase the OD untruthfully. It's better to wash the wells with PBS before adding the MTT.
hi
you should check the expire date of drug and drug potency before the use .
Article Antiproliferative Activity of the Chinese Medicinal Compound...
Article Wu Q, Wang R, Yang Q, Hou X, Chen S, Hou Y, Chen C, Yang Y, ...
good luck
Some cell respond to some cytotoxic drugs in the way you described. E.g., in our hands, MCF7 cells give a "plato" at appr. 50% in quite wide range of concentrations of some platinum comlexes, so, no one can say which particular concentration should be considered as IC50.
Sometimes, when IC50 is absolutely required, you have to change your model (I mean choose another cell line).
IC50 values described in the literature could be helpfull to some extent, but you have to find it for your particular lab.
As it was mentioned by Reza Hadavi, your drug could be out of date. Unfortunately, cells can change too (the more passages, the more changes); and your HepG2 could easily be something different from what we can see at atcc.org. So, if your HepG2 cell line differs considerably from thous described in literature, buying new vial of HepG2 from a trusted source like atcc might be an option.
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