An Exploration Research Of Colorectal Normal-adenoma-adenocarcinoma Sequence

BackgroundColorectal cancer(CRC)accounts for 9.8%of all annually diagnosed cancers(3rd rank)and for 9.2%of cancer-related deaths(2nd rank).The most canonical carcinogenesis pathway is "normal-adenoma-adenocarcinoma" sequence accounting for 85%of all CRCs.Therefore,in-depth understanding of this evolution process and exploring the key proteins that affect this evolution process will help to carry out target prevention of colorectal cancer.However,there is currently a lack of systematic in-depth analysis of the evolution process of normal-adenoma-adenocarcinoma sequence at the protein level.Aim(1)Exploring the protein enrichment changes in the evolution of normal-adenoma-adenocarcinoma sequence;(2)Exploring the key proteins in the evolution of normal-adenoma-adenocarcinoma sequence and the relationship between these proteins and the prognosis of patients with colorectal cancer;(3)Exploring the protein expression characteristics of tubulovillous(TVA)and the representative protein of potential malignant characteristics of TVA;(4)Exploring the key proteins in the evolution of high-grade intraepithelial neoplasia-adenocarcinoma.Methods(1)We used in-depth quantitative proteomics technology to perform protein profiling experiments on normal epithelial tissues,adenoma tissues and adenocarcinoma tissues,obtain protein expression profiles of each tissue,screen differentially expressed proteins,and used GSEA and Proteomaps methods to analyze this evolution Process protein enrichment analysis;(2)The key proteins in normal-adenoma-carcinoma sequence were found by combining with the external transcriptomic database and proteomic database,following the"identification-verification-validation" process,and constructing protein-protein interaction networks.We used the internal database to evaluate the relationship between key proteins and the prognosis of colorectal cancer patients.Furthermore,we performed functional experiments at the level of adenoma organoids and tumor cell lines to confirm the promotion effect of key proteins in this evolutionary process;(3)We constructed heatmaps and combined external transcriptomic and proteomic databases for protein enrichment analysis by bioinformatics tools.and performing Functional experiments of the representative proteins of TVA's potential malignant features were performed at the adenoma organoid level;(4)The key proteins in the evolution process of high-grade intraepithelial neoplasia-adenocarcinoma were identified by the integrating analysis of internal proteomics database,external transcriptomics database and proteomics database.Results(1)The protein expression profile changes in the process of normal-adenoma are mainly enriched in the ribosome and spliceosome-related pathways.The core protein in this evolution is RRP12;(2)The protein expression profile changes in the process of adenoma-adenocarcinoma are mainly enriched in the extracellular matrix and epithelial-mesenchymal transition related pathways.The core protein of this evolution process is SERPINH1;(3)SERPINH1 not only plays an important role in the progression of adenoma to adenocarcinoma,but also promotes the development of colorectal cancer.It is related to the epithelial-mesenchymal transition pathway and is an independent risk factor for the prognosis of colorectal cancer patients;(4)The typical protein expression characteristics of TVA are related to mitochondria and metabolic reprogramming pathways,such as oxidative phosphorylation,branched chain amino acid degradation and mitochondrial ribosomal pathway;the representative protein of potential malignant features of TVA is PLOD3;PLOD3 is related to the "immune desert" phenotype in the tumor microenvironment of colorectal cancer,and is a potential prognostic marker for CRC patients;(5)The protein expression profile changes in the evolution of high-grade intraepithelial neoplasia to adenocarcinoma are mainly enriched in extracellular matrix tissue,collagen synthesis,defensive response to bacteria and fungi,neutrophil degranulation and cell proliferation.Pathway or biological process;SERPINH1 and P3H1 are the key proteins in this evolutionary process;SERPINH1 and P3H1 are related to the consensus molecular subtype of mesenchymal colorectal cancer,and are related to the immunosuppressive phenotype in the tumor microenvironment.ConclusionBased on quantitative proteomics technology,we have conducted an in-depth and comprehensive exploration of the evolution process of normal-adenoma-adenocarcinoma sequence.RRP 12 and SERPINH1 are the key proteins in the evolution of normal-adenoma and adenoma-adenocarcinoma process,respectively.SERPINH1 can promote the progression of colorectal cancer and is also one of the key proteins in the evolution of high-grade intraepithelial neoplasia-adenocarcinoma.PLOD3 is a representative protein of the malignant characteristics of TVA.P3H1 and SERPINH1 are key proteins in the evolution of high-grade intraepithelial neoplasia to adenocarcinoma.The above proteins may be potential targets for the targeted prevention of colorectal cancer.