Abnormal ECA-Binding Membrane Glycans and Galactosylated CAT and P4HB in Lesion Tissues as Potential Biomarkers for Hepatocellular Carcinoma Diagnosis

Abstract

Hepatocellular carcinoma (HCC) is one of the most common types of cancer. Despite decades of research efforts, the search for novel biomarkers is still urgently needed for the diagnosis of HCC and the improvement of clinical outcomes. Previous studies of HCC clinical biomarkers have usually focused on serum and urine samples (e.g., serum Alpha-fetoprotein (AFP). However, cellular membrane proteins in lesion tissues are less used in HCC diagnosis. The abnormal expression of membrane glycoproteins in tumor lesions are considered as potential targets for tumor diagnosis and tumor therapies. Here, a lectin array has been employed to screen and identify abnormal glycopatterns and cellular membrane glycans in HCC lesion tissues compared with adjacent non-tumor tissues. We found that there was significantly less expression of Erythrina cristagalli (ECA) lectin binding (Galβ1-3/β1-4) glycans on the cellular membrane of HCC lesion tissues compared with those of adjacent non-tumor tissues. Immunohistochemistry analysis further showed that ECA-binding ability on the membrane proteins of HCC tissues progressively decreased in different tumor-node-metastasis (TNM) stages (stage I to stage III) as the malignancy of liver cancer increased. Receiver operating curve (ROC) analysis showed ECA-binding ability yielding a sensitivity of 85% and specificity of 75%, and a combination of ECA and AFP has better clinical diagnostic efficiency, yielding a sensitivity of 90% and specificity of 85%, than ECA or AFP assay alone. ECA pull-down followed by mass spectrometry further showed that there was significantly less expression of ECA binding membrane catalase (CAT) and prolyl 4-hydroxylase beta polypeptide (P4HB) in HCC tissues compared with the adjacent non-tumor tissues. The abnormally increased expression of total CAT and P4HB and decreased expression of galactosylated membrane CAT and P4HB in HCC cell lines were correlated with an HCC metastasis status. Our findings suggest that abnormal declined ECA-binding galatosylated membrane glycans and two galactosylated-CAT and P4HB glycoproteins in lesion tissues are potential biomarkers in the diagnosis and/or metastasis prediction for HCC.

MiR-204-5p regulates HUVEC cell inflammation and apoptosis by targeting P4HB

“…Assay data from clinical samples also supported the remarkably high expression of P4HB in the serum of DVT patients. Recently, P4HB was recognized to be a novel diagnostic and prognostic biomarker in several malignancies (24,36). Differential protein analysis of the midbrain ischemia/reperfusion rat cerebral cortex based on i-TRAQ quantitative proteomic probing reveals that P4HB exerts a key role in oxidative stress, apoptosis and inflammatory responses (37).…”

Targeting Prolyl 4-Hydroxylase Subunit Beta (P4HB) in Cancer: New Roads to Travel

“…In hepatocellular carcinoma (HCC), P4HB was overexpressed in HCC tissues and cell lines and higher P4HB levels were associated with advanced disease and worse prognosis [48,49]. In vivo experiments, the overexpression of P4HB could promote HCC cell growth, migration, invasion and epithelial-to-mesenchymal transition (EMT), while P4HB knockdown could inhibit HCC tumorigenesis [48,50].…”

An integrated study to decipher immunosuppressive cellular communication in the PDAC environment.

“…[21] P4HB was found to be overexpressed in various cancer types including bladder cancer, renal clear cell carcinoma, hepatocellular carcinoma, and glioblastoma. [24][25][26][27] However, several microarray studies showed that P4HB is downregulated in PDAC. [28][29][30] On the other hand, the expression of this gene was indicated to be related with the stage of the cancer; in the first and fourth stages it is downregulated while upregulation is reported in the second and third stages of PDAC.…”