study of receptors in cell lines - (Mar/15/2013 )
I'm styudying receptor in cancer cell lines. Since it is overexpressed I plan to observe the effect of a receptor antagonist in the cancer cell lines (e.g. MTT by proliferation). My problem is that the antagonist that's available may not be 100% selective to the recetor I'm studying (e.g receptor A) and that could also affect receptor B as well.
How do I ensure that the B receptor -selective antagonists are not active in my assay to have more compelling results.
I was thinking I treat the cell lines with antagonist to receptor A alone, antagonist to receptor B alone and combination of both antagonists of A and B?
its true no antagonists can be 100% selective, it does have some known or unknown off-targets, so may be first you got to optimize the minimum drug concentration to get a max negative effect of the prime target and less antagonistic effect on the suspected off target. you can ensure this by analysing the receptor phosphorylation and/or their substrate activation if they have any specific non-overlaping susbtrates (here the chances are very dim though!!!)...
after this you can go into the functional assays dealing prolfieration, survival, etc etc....,based on the biological activity of your target receptor....