| Objective:Colorectal cancer(CRC)is the third most commonly diagnosed cancer worldwide.Thus,there is an urgent need for novel and more specific biomarkers to improve early diagnosis and patient outcome.To investigate the mechanism and the biological function of proteins in CRC progression,a shotgun proteomics were performed to identify novel biomarkers involved in CRC.Methods:In this study,CRC and matched normal mucosal tissue proteome profiles were performed by label-free mass spectrometry(MS)to identify novel target involved in CRC.The differential expressed proteins were validated by western blot and immunohistochemistry.Using Real-Time Cell Analyzer and total internal reflection fluorescence microscopy,the role of glucagon was studied in carcinogenesis of colorectal cancer.Results:In six pairs of colorectal cancer tissue,in total 169 differentially expressed proteins were identified.ANXA4,PGK,and RAN,were validated as being upregulated in six CRC tissues with a 1.95,2.27 and 2.22-fold change.The expression of ATP5A,CALR,CAMKII and Glucagon were significantly decreased in CRC with a 2.00,1.76,1.84 and 1.92-fold change(P<0.05).Moreover,glucagon inhibited the proliferation and migration of CRC cells by using real time cell analysis.Added glucagon on colorectal cancer cell,glucagon inhibited the proliferation of colorectal cancer cells by 2.1-fold(SW480)and 1.4-fold(CT26)compared to the control group.The migration was 1.4-fold(SW480)，and 1.9-fold CT26.)inhibited after 50 hours.Adenylate cyclase inhibitor(2’5’dideoxyadenosine,DDA)reduced inhibitory effect of glucagon on colorectal cancer cells.It was shown the similiar effect of sarcoendoplasmic reticulum Ca2+-ATPase(SERCA)inhibitor CPA.Conclusion:This demonstrates that glucagon is involved in the cAMP/Ca2+ signal transduction pathway to inhibit the proliferation and migration of colorectal cancer cells in CRCs progression and thus could be potentially used as a new target for the treatment of CRCs. |
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