The Abnormal Glycopatterns of Salivary Glycoproteins in Esophageal Squamous Cell Carcinoma Patients

Abstract

Glycosylation is one of the most crucial posttranslational modifications of proteins, containing a remarkable amount of biological information. The alteration of glycosylation is closely associated with certain diseases. Exploring glyco-code in the development of diseases is a hot topic in recent years. Esophageal squamous cell carcinoma (ESCC) is the primary pathological histology in developing countries and a severe threat to human health. Although the glycan profiles in the blood samples of ESCC patients were analyzed using glycomic and glycoproteomic methods, the difference of salivary glycopatterns between healthy subjects and ESCC patients is not explicit yet. In the present study, ESCC patients (n = 16) and healthy volunteers (HVs, n = 25) were enrolled. The glycomic strategy combining lectin microarray and lectin blotting was employed to investigate and confirm the altered salivary glycopatterns. Datura stramonium (DSA) was selected to isolate the GlcNAc or Galβ1-4GlcNA-containing glycoproteins due to the distinct difference between ESCC patients and HVs. The N-glycans from DSA-enriched glycoproteins were released by PNGase F and further identified by MALDI-TOF/TOF-MS to obtain the precise structural information of the altered glycans. As a result, the glycopatterns recognized by 13 lectins (e.g., ECA, RCA120, and DSA) showed significant alterations in ESCC patients’ saliva. The ESCC patients showed higher levels of GalNAc and Gal, sialic acid, and GlcNAc expression profiles and lower levels of mannose and fucose expression profiles. The MALDI-TOF/TOF-MS results indicated that the proportion of the GlcNAc or Galβ1-4GlcNAc-containing N-glycans was increased in ESCC patients (79.04%) compared with HV (63.20%), which was consistent with the results of lectin microarrays. Our findings provide comprehensive information to understand the complex physiological changes in ESCC patients. And the altered salivary glycopatterns such as GlcNAc or Galβ1-4GlcNAc-containing N-glycans recognized by DSA might serve as potential biomarkers for the diagnosis of ESCC patients.

N-Glycan Profiles of Neuraminidase from Avian Influenza Viruses

“…Similar to our previous reports [25,26], the N-glycans of the isolated NAs were released by PNGase F glycosidase. The purified N-glycans of NA were characterized by MALDI-TOF/TOF-MS (Ultra eXtreme, Bruker Daltonics; Bremen, Germany) as described previously.…”

Glycan characteristics of human heart constituent cells maintaining organ function: relatively stable glycan profiles in cellular senescence

“…A better understanding of each cellular condition is needed for advancing research on regenerative therapy due to differences in the various factors, including diseases and ages. In addition, further analysis of glycan composition via methods such as mass spectrometry will enhance understanding when combined with analysis of glycan structures, including branching, coordination, and binding modes (Shu et al 2021 ). Investigating optimal cellular ratios for remodeling and the carrier proteins of characteristic glycans will be important in making these glycans useful as indicators for clarifying the details of personalized cardiovascular conditions and cellular functions in patients.…”

Latest insights into the global epidemiological features, screening, early diagnosis and prognosis prediction of esophageal squamous cell carcinoma

“…N-linked glycoproteomic profiling analysis indicated that the expression of total procathepsin D and high-mannose procathepsin D was upregulated, while the expression of total haptoglobin, high-mannose clusterin and the GlcNAc/sialic acid-containing fraction of 14-3-3ζ was downregulated in ESCC tissues[ 91 ]. In addition, Shu et al [ 92 ] reported that the proportion of ESCC patients with GlcNAc or Galβ1-4GlcNAc-containing N-glycans was increased. Similarly, the expression of glycosylated proteins was distinct between the saliva of ESCC patients and that of healthy controls, and screening for ESCC biomarkers in saliva is noninvasive and may be a more acceptable approach[ 92 ].…”

“…In addition, Shu et al [ 92 ] reported that the proportion of ESCC patients with GlcNAc or Galβ1-4GlcNAc-containing N-glycans was increased. Similarly, the expression of glycosylated proteins was distinct between the saliva of ESCC patients and that of healthy controls, and screening for ESCC biomarkers in saliva is noninvasive and may be a more acceptable approach[ 92 ]. In addition to screening and early diagnosis, glycosylated proteins may also be associated with the metastasis and prognosis of ESCC.…”