High-throughput screening of tumor metastatic-related differential glycoprotein in hepatocellular carcinoma by iTRAQ combines lectin-related techniques

Abstract

Glycoproteomics is an important aspect in the research of cancer biomarker discovery. The objective of our study is to screen the profile of serum glycoproteins in hepatocellular carcinoma (HCC) patients and to discover differentially expressed glycoproteins in HCC with or without metastasis. We collected serum from HCC patients and divided them into two groups (non-metastatic HCC group and metastatic HCC group) according to 2002 UICC TNM staging system. Wheat germ agglutinin (WGA) lectin was used to enrich the serum glycoproteins by lectin affinity chromatography. The enriched glycoproteins were labeled with mass-balanced isobaric tags (iTRAQ) and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Two differential glycoproteins were validated by Western blot and biochemical methods, respectively. Fifteen differential serum glycoproteins with WGA affinity were identified (p < 0.05). Among them, nine proteins were up-regulated (>1.5-folds) and six were down-regulated (<0.5-folds) in HCC patients with metastasis. Expression of alpha-1-antitrypsin (SERPINA1) and apolipoprotein A-I (APOA1) was validated by Western blot and biochemical methods, respectively (p < 0.05). Our study has obtained a set of HCC metastasis-associated glycoproteins which may serve as novel prognostic candidates and potential therapeutic targets for HCC metastasis. SERPINA1 might act as a potential glycoprotein biomarker of HCC metastasis.

LC–MS/MS Quantitation of Esophagus Disease Blood Serum Glycoproteins by Enrichment with Hydrazide Chemistry and Lectin Affinity Chromatography

“…21 It is a circulating serine protease inhibitor or serpin and an acute-phase protein secreted in the liver. It is involved in the inhibition of apoptosis, or modulation of local and systemic inflammatory responses.…”

“…These two enrichment techniques have been successfully employed in many studies of glycoproteins associated with various cancer types including lung cancer, 12−15 breast cancer, 16−18 prostate cancer, 19,20 and hepatocellular carcinoma. 21−23 …”

Quantitative proteomics analysis of early recurrence/metastasis of huge hepatocellular carcinoma following radical resection

“…So far, several groups have reported the applying of iTRAQ based quantitative proteomics approach in the study of hepatic cancer, especially for the screening of diagnostic or prognostic protein biomarkers. He et al have reported the serum biomarker screening of AFP negative HBV related HCC through the iTRAQ based approach [35]; Ko et al have reported the iTRAQ based quantitative analysis of HCC cancer stem cell proteome [36]; Huang et al have reported the iTRAQ based serum biomarker screening of the HCC micro-vascular invasion [37]; Qin et al, Yu et al and Wang et al have reported the screening of metastatic related proteins of HCC through iTRAQ based approach [33,38,39]; the iTRAQ based quantitative study of proteome change during HBV infection has also been reported [40,41]. However, the application of iTRAQ labeling in studying the molecular mechanisms and screening for biomarkers associated with the early recurrence/metastasis of H-HCC has not yet been reported to our knowledge.…”

Preclinical Evaluation of Transcriptional Targeting Strategy for Human Hepatocellular Carcinoma in an Orthotopic Xenograft Mouse Model

“…While the mechanism responsible for its complex hepatocarcinogenesis is still under investigation, rapid hepatocellular carcinoma tumor proliferation due to dysregulation of the cell-cycle proteins is clear. Herein, we have taken advantage of the rapid proliferating nature of tumor cells in general and the high levels of hAAT expression found in hepatocellular carcinoma (30)(31)(32) and generated an effective vector system whereby transgene expression is dependent on the activation of two endogenously regulated promoters, i.e., 4ÂApoE/hAAT liver-specific promoter and recombinant human cyclin A2. In the context of oncolytic HSV-1, the 4ÂApoE/hAAT liver-specific promoter has also been shown to drive the expression of complementary sequences of selected miRNAs exclusively in hepatocellular carcinoma (33).…”