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

Ying Kong; Hao Chen; Mengyu Chen; Yongshuai Li; Jiarong Li; Qi Liu; Huan Xiong; Tangxi Guo; Yan Xie; Yufeng Yuan; Xiao-Lian Zhang

doi:10.3389/fonc.2022.855952

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

Front Oncol. 2022 Mar 22:12:855952. doi: 10.3389/fonc.2022.855952. eCollection 2022.

Authors

Ying Kong¹, Hao Chen², Mengyu Chen¹, Yongshuai Li¹, Jiarong Li¹, Qi Liu¹, Huan Xiong¹, Tangxi Guo³, Yan Xie¹, Yufeng Yuan⁴, Xiao-Lian Zhang^{1

5}

Affiliations

¹ Hubei Province Key Laboratory of Allergy and Immunology, and Department of Immunology, Wuhan University School of Basic Medical Sciences, Wuhan, China.
² Department of Pathology, Zhongnan Hospital of Wuhan University, Wuhan, China.
³ Department of Radiation and Medical Oncology, Zhongnan Hospital of Wuhan University, Wuhan, China.
⁴ Hepatobiliary and Pancreatic Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.
⁵ Allergy Department of Zhongnan Hospital, State Key Laboratory of Virology, Medical Research Institute Wuhan University School of Medicine, Wuhan, China.

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.

Keywords: Erythrina cristagalli (ECA) lectin; catalase (CAT); hepatocellular carcinoma; lectin array; membrane biomarker for diagnosis; prolyl 4-hydroxylase beta polypeptide (P4HB).