| RNA sequencing(RNA-seq)technology has been widely used in the field of life science.The emergence of single-cell RNA sequencing(scRNA-seq)technology has revolutionized the research methods in this field.Owing to its unique advantages such as exploring cellular heterogeneity,identifying cell subtypes,figuring out cell developmental process and so on,scRNA-seq technology has gradually become a powerful tool for in-depth study of complex transcriptomes,and is often used in medical related fields such as immunology and oncology.Colorectal cancer(CRC)is one of the most common malignant tumor diseases in modern society.The transcriptomic research of CRC is of great significance for the disease’s diagnosis and treatment.Some questions such as cellular heterogeneity in CRC are unclear due to low cell flux in earlier research on CRC using scRNA-seq technology.Therefore,we used 10 x Genomics technology to deal with the tumor tissue samples from two colorectal cancer(CRC)patients and the control intestinal tissue sample from one healthy donor and explore cellular heterogeneity in colorectal cancer through bioinformatic analysis.In terms of bioinformatics analysis,we summarized the main processes of scRNA-seq data analysis: using Cell Ranger software for data quality control and sequence alignment;using R software packages Seurat and cluster Profiler for cell and gene screening,cell type identification,differential gene analysis and their enrichment analysis;using Velocyto software for cell trajectory inference.In addition,we paid special attention to the processing of doublets in scRNA-seq data,and built an analysis process based on R to identify them.We conducted data analysis through the above processes we concluded.We identified 8 global cell types and 6epithelial cell subtypes in 3 samples and found disease-associated signaling pathways through differential gene enrichment analysis,which is of great significance for the clinical diagnosis and prognostic detection of colorectal cancer. |
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