Recently I ran a Microarray analysis on cells treated with purified plant-extracted compound, however, the caspase 3 levels were elevated after 24 hours suggesting high apoptotic activity of treatment but the fold change decreased after 48 hours of treatment given the fact that the cell count dropped drastically after 48 hours and the yielded RNA was way less then the one yielded at 24 hours? another fact that the P21 expression was very high at 48 hours.
I assumed that after 48 hours cells were undergoing apoptosis but the expression levels were low due to low cell count, so far I couldn't find papers to support this finding.
another assumption is that the cells that managed to proliferate and continue to grow after 24 hours were in a different cell cycle phases ( mother & daughter cells) which might have contributed to drug- resistance, or just the 24 hours exposure wasn't enough for all the cell to diminish but a longer exposure was required with the dosage used.
would you please help me? ( I feel lost)
your assumption 2 may be correct
cell synchronous required
also cell number matters.
p21 is proliferation marker if enhances it means cells are growing fast while caspase is apoptotic marker.
caspase can play dual role in implantation its expression is different whereas in cancer it is only proliferative marker like PTEN.
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