Recently I got a unchanged qRT-PCR result of one gene and significantly changed target protein quantified by western-blot, if both experiments are true, how to explain it?
Hello Jingtao.
That would be called post-transcriptional regulation, not to mention about miRNA regulation to break mRNA.
There is an issue that PCR amplify greatly the real-expressed genes, making bigger or smaller gaps.
Regards.
Relationship between transcript and protein is not always consistant for variety of reasons. Hence why validation is necessary.
https://www.nature.com/articles/srep10775
Article Maier T, Güell M, Serrano L.. Correlation of mRNA and protei...
Thank you guys. Even the target protein level significantly increased precisely quantified by western-blot, it does not necessarily prove that protein/enzyme activity is enhanced, so why bother measuring mRNA or protein, the real aspect need to concern is the last step protein /enzyme activity, which though sometimes is hard to measure.
How often have you seen a case where protein expression was induced but not the protein activity? It could happen but in my experience this is minimal. It might be much easier to fine-tune protein activity for the cell by means of gene regulation than by say post-translational modification i.e. by phosphorylation which can require complex cascade of events to trigger. In addition, you also have phenotypic/functional indicators which basically should correlate with your hypothesis, i.e. an proto-oncogene is induced in expression, therefore there should be more cell growth, etc. Not all proteins are enzymes, and by all means if you can measure the activity of a protein by a method i.e. by imaging that further strengthens your hypothesis and methodology.
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