Glycopattern Alteration of Glycoproteins in Gastrointestinal Cancer Cell Lines and Their Cell-Derived Exosomes

Abstract

Gastrointestinal (GI) cancers constitute the largest portion of all human cancers, and the most prevalent GI cancers in China are colorectal cancer (CRC), gastric cancer (GC), and hepatocellular carcinoma (HCC). Exosomes are nanosized vesicles containing proteins, lipids, glycans, and nucleic acid, which play important roles in the tumor microenvironment and progression. Aberrant glycosylation is closely associated with GI cancers; however, little is known about the glycopattern of the exosomes from GI cancer cells. In this study, glycopatterns of HCC, CRC, and GC cell lines and their exosomes were detected using lectin microarrays. For all exosomes, (GlcNAcβ1-4) n and Galβ1-4GlcNAc (DSA) were the most abundant glycans, but αGalNAc and αGal (GSL-II and SBA) were the least. Different cancers had various characteristic glycans in either cells or exosomes. Glycans altered in cell-derived exosomes were not always consistent with the host cells in the same cancer. However, Fucα1-6GlcNAc (core fucose) and Fucα1-3­(Galβ1-4)­GlcNAc (AAL) were altered consistently in cells and exosomes although they were decreased in HCC and CRC but increased in GC. The study drew the full-scale glycan fingerprint of cells and exosomes related to GI cancer, which may provide useful information for finding specific biomarkers and exploring the underlying mechanism of glycosylation in exosomes.

Biotin–Avidin System-Based Delivery Enhances the Therapeutic Performance of MSC-Derived Exosomes

“…Exosome experiments were conducted after cleaning the sample probe, as previously described. 79 According to previously published studies that performed exosome analysis, 80,81 Cellular Uptake Assays. The PKH26 red fluorescence labeling kit (Ex 551 nm/Em 567 nm; Sigma) was used to label the obtained and purified MSC-EXs according to the instructions provided with the kit.…”

“…Exosome experiments were conducted after cleaning the sample probe, as previously described . According to previously published studies that performed exosome analysis, , the threshold in violet side scattering height (VSSC-H) was manually set to 1500 to block background events, and the acquisition setting in the VSSC-H gain signal was set to 10. To increase the accuracy, the flow speed was adjusted to 10 μL/min, the concentration was adjusted to less than 10,000 particles/μL by appropriate dilution of particle-free PBS during the measurements, and 100000 events were ultimately acquired.…”

Lectins as potential tools for cancer biomarker discovery from extracellular vesicles

“…Recently, blotting with six lectins was done in a study that reported the presence of disease-associated glyco-epitopes in bladder cancer-derived EVs [ 184 ]. In another study, after 20-lectin microarray identified altered glycans on EVs from gastrointestinal cancer, findings were validated using lectin blotting [ 185 ].…”

Detecting early‐warning biomarkers associated with heart‐exosome genetic‐signature for acute myocardial infarction: A source‐tracking study of exosome

“…Exosomes (measuring 30–150 nm in diameter), which are lipid belayer‐enclosed vesicles, are released by living cells into the extracellular space in response to hypoxia and inflammation. 2 , 3 They act as intracellular messengers, plasma through all body fluids, and carry a range of biologically active cargo, including proteins and genetic materials, from the host cells. This has been shown to indicate pathophysiology or physiological changes and has emerged as a non‐invasive liquid biopsy strategy for cardiovascular diseases.…”