Proteomic analysis of muscarinic acetylcholine receptor-mediated proliferation in HT-29 human colon cancer cells

Muscarinic acetylcholine receptors (mAChRs) are members of G-protein-coupled receptors. They can induce agonist-dependent neoplastic transformation and facilitate colon cancer proliferation via promoting rapid expression of a variety of early responsive genes. In this study, we used 2-dimensional gel electrophoresis (2-DE) approach with subsequent mass spectrometry (MS) to identify up- and down-regulated proteins (a total of 23 protein spots) involved in mAChRs-related signaling pathway, energy metabolism, transcription/translation, oxidative stress metabolism and cytoskeleton organization in agonist carbachol stimulated HT-29 human colon cells. We found that the increased expression of adenocarcinoma biomarker, annexin A5 (ANXA5) induced by carbachol treatment, which was confirmed by immunoblot. This study contributes to the understanding of mechanisms underlying mAChRs agonist-induced expression of whole proteins in HT-29 colon cancer cells. Our results indicated that ANXA5 might serve as a potential biomarker for the diagnosis of colon cancer.