Majority of the studies have shown that MCF-7 cells do not express caspase-3, which is however the major apoptotic pathway for a wide range of cancer cells. And if different drugs activate different caspase pathways, which caspase activity should I look for in caspase-3 deficient MCF-7 cells?
Other caspases such as caspase 6, 7, 8, or 9 could participate in apoptosis. Therefore, using inhibitors that inhibit these caspases specifically could identify a candidate caspase(s) in MCF-7 cells.
MCF -7 cells do not express caspase-3. However, these cells do undergo mitochondria mediated apoptosis in which caspase-7 is involved. You can use DEVD peptide substrate which is a substrate for both caspase-3 and caspase-7.
You can run a western an look for caspase 2, caspase 6 and 7 even 8 to see if this ones are activated (cleaved). However, this paper shows that is caspase 9, 6 and 7 that can be activated for apoptosis in MCF7. There is a very clean western for caspase 7
http://www.nature.com/onc/journal/v20/n45/pdf/1204815a.pdf
Good luck
MCF-7 cells have a truncated CASP3 that is not fully transcribed. In some of our studies, the truncated mRNA is picked up by Affymetrix microarrays and you can see its transcriptional regulation. However, there is no full length transcript and no translated protein for CASP3. Many of the other caspases are expressed and appropriately regulated. CASP7 appears to function in lieu of the lack of CASP3 (or at least that seems to be the prevailing sentiment in the literature; its probably a little more complicated than that). CASP7, CASP8 and CASP9 are regulated by interferons and affect responsiveness to antiestrogens (if I remember correctly, CASP1 (ICE) is also regulated in MCF-7 cells and variants). You can read what we found in several of our papers but the most useful might be Ning et al., Mol Cancer Ther 9: 1274-85, 2010. It should be accessible here on Research Gate. If you want a copy and cant find it, let me know and I'll send it.
I hope this is helpful...Bob
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