Table 3.

Summary the major effect of the studies describing NETs in CRC

Study design	Cell type	Potential mechanisms to induce NET formation	The role of NET in tumor progression	Frist author, References
In vivo and in vitro	Murine colorectal (MC38) cells, HCT116, Hepa1-6, and Huh7 cell lines	Increased citrullinated histones and circulating MPO-DNA levels were related to poor survival of CRC patients	NETs can directly alter the metabolic programming of cancer cells to increase tumor growth and shorter survival time	Yazdani et al. [129]
In vivo, in vitro nd ex vivo	Human colorectal cell line HCT116 or luciferase-labeled HCT116 cells	TLR9 and the mitogen-activated protein kinase signaling pathway	LPS-induced formation of NETs in promoting the development of tumors and metastasis	Wang et al. [130]
In vitro and Ex vivo	Human acute myeloid leukemia (AML) cells, Caco-2 cells	–	Confirmed presence of NETs within the primary tumor sites of CRC and gradually dispersed to the tumor boundary, particularly to nearby metastatic lymph nodes	Arelaki et al. [122]
In vivo and in vitro	DKs-8(WT allele) cells and DKO-1 (KRASmutant)cells	Exoxsomes from KRAS mutant CRC increase IL-8 production and provoke NET formation	Released NETs increase CRC cells growth	Shang et al. [131]
In vivo, in vitro,and Ex vivo	Human hepatocellular carcinoma, human cell line HT29, mice cell line MC38	Elevated tumorous interleukin (IL)-8 expression triggered by NETs and overproduced IL-8 in turn activate neutrophils towards NETs formation	Increased NETs boosted tumorous proliferation and invasion and contributed to onset of CRC liver metastasis	Yang et al. [132]
In vivo and in vitro	Human colon carcinoma cell line (HT-29), murine colon carcinoma subline with low CEACAM1 expression (MC38CC1-),murine colon carcinoma subline stably transfected with CEACAM1 long isoform[MC38CC1L]	NET-associated carcinoembryonic Ag cell adhesion molecule 1 (CEACAM1) as an essential element for this interaction	NETs can promote colon carcinoma cell adhesion, migration and metastasis	Rayes et al. [133]
In vivo and in vitro	Murine Lewis Lung carcinoma cell subline H59, Murine colon carcinoma cell line MC38	Primary colon cancer cells provoked NETs generation	Prime adhesion of CTCs to the liver and degradation of NETs decreased CRC cell adhesion and spontaneous metastasis to the liver and lung	Rayes et al. [118]
In vivo and Ex vivo	human colon cancer cell line HCT116,	The transmembrane protein CCDC25 as a NET-DNA receptor on cancer cells that senses extracellular DNA and subsequently activates the ILK-β-parvin pathway to enhance cell motility	A transmembrane DNA receptor that mediates NET-dependent metastasis	Yang et al. [136]
In vivo and in vitro	Human hepatoma cell line HepG2, murine colon carcinoma MC38	Neutrophil infiltration and NET formation reduced by adeno-associated virus (AAV) based DNase I gene therapy	Reduced liver metastasis	Xia et al. [137]
Ex vivo	Human umbilical vein endothelial cells (HUVECs)	platelets from CRC patients stimulated healthy neutrophils to extrude NETs, which could be inhibited by the depletion of HMGB1	NETs induce the procoagulant activity PCA and promote hypercoagulable state in CRC	Zhang et al. [142] Guglietta et al. [139]
In vivo, in vitro,and Ex vivo	MC38 and Luciferase-expressing MC38 cells (MC38/Luc)	NET triggered HMGB1 release and activated TLR9-dependent pathways	NETs further fuel cancer cells adhesion, proliferation, migration, and invasionthe and reduce more than fourfold disease free survival	Tohme et al. [128]
Ex Vitro	Systemic neutrophils were isolated from human		Adverse patient outcomes were associated with increased preoperative NETs production	Richardson et al. [142, 143]