Hello everyone.
I am looking at studying the connection between NF-kB and JAK/STAT3 in normal and cancer cells specifically how chronic inflammation leads to development of colorectal cancer. I would like to test how the levels of IL-6 affect both STAT3 and NF-kB expression.
Does anyone have any advice on how they would test this? Including use of controls and additional cytokines, and/or upstream/downstream targets of STAT3 and NF-kB
I am having trouble putting together this experiment so any help would be greatly appreciated. Thank you very much.
I think you might need to have a compound or drugs that could induce inhibition for one of your targets genes such as NF-KB. From there you can observed the effect of NF-KB on JAK/STAT3 and IL6 for instance. You may conduct real-time PCR to evaluate the relation of those genes. Plus, you may also try western blot as it will give you a strong results.
Dear Connor,
The subject of your study is interesting. In case of inflammation and development of colorectal cancer, we conducted a series of experiments using both animal models and in vitro approaches. We found a significant upregulation in Toll-like receptor 2 and 4 as well as MyD88 as main upstream regulators of both NF-kB and IL-6 in intestinal tissue. Using IL-6 antibody would be helpful. However, I suggest you to read the following review that may help you.
Waldner, Maximilian J., and Markus F. Neurath. "Master regulator of intestinal disease: IL-6 in chronic inflammation and cancer development." Seminars in immunology. Vol. 26. No. 1. Academic Press, 2014.
Good luck
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