A preliminary analysis of transcriptome expression differences and functions of colorectal cancer peripheral blood DNT cells based on SMART-seq2 sequencing
10.13431/j.cnki.immunol.j.20230117
- VernacularTitle:基于SMART-seq2测序分析结直肠癌外周血DNT细胞转录组的表达差异及功能初探
- Author:
Yuling DAI
1
;
Yunbo WEI
;
Jing WU
;
Huiru ZHU
;
Xiaofei LIU
Author Information
1. 261053,潍坊医学院医学检验学院
- Keywords:
Colorectal cancer;
Double-negative T cells;
Differential gene;
SMART-seq2
- From:
Immunological Journal
2023;39(10):900-909
- CountryChina
- Language:Chinese
-
Abstract:
This study was performed to explore the differential genes and functions of double-negative T cells in peripheral blood of patients with colorectal cancer.Two colorectal cancer patients and two healthy physical examiners were selected,and peripheral blood double-negative T cells were firstly sorted by flow cytometry,and then sequencing data were obtained using single cell full-length transcriptome(SMART-seq2)sequencing technology to screen differentially expressed genes.The screened differentially expressed genes were subjected to Gene Ontology Enrichment(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.The STRING database was used to construct the protein interaction network and identify key genes by Cytoscape software;RT-qPCR was used to verify the differential expression genes in DNT cells.Compared with healthy subjects,there were 1 276 peripheral blood double-negative T-cell differential genes in colorectal cancer patients,including 141 up-regulated genes and 1 135 down-regulated genes.GO analysis showed that the differential genes were mainly involved in biological functions such as methylation,metabolic processes and transferase activity;KEGG pathway analysis showed that the differential genes were mainly involved in signaling pathways such as autophagy,P53 signaling pathway and phosphatidylinositol metabolism.The protein interaction network contains 1 154 nodes and 1 022 edges,in addition,10 hub genes were identified:PIK3C3,WIPI1,ATG101,PIK3R4,DDX10,RBM28,SDAD1,ATG16L1,UVRAG,ATG7.RT-qPCR validated 10 differentially expressed genes,of which 7 differentially expressed genes showed trends consistent with sequencing results,and 3 genes showed expression inconsistent with sequencing results.DNT cells may be involved in the development of colorectal cancer through methylation,P53 signaling pathway and autophagy,and at the same time,DNT cells may inhibit the development of colorectal cancer through the regulation of genes.This study provides a theoretical basis for further investigation of the function of DNT cells in malignant tumors.
