Quantitative Proteomics Analysis of Tissue Interstitial Fluid for Identification of Novel Serum Candidate Diagnostic Marker for Hepatocellular Carcinoma

Abstract

Hepatocellular carcinoma (HCC) is the fifth most common malignant cancer in the world. The sensitivity of alpha-fetoprotein (AFP) is still inadequate for HCC diagnosis. Tissue interstitial fluid (TIF), as the liquid microenvironment of cancer cells, was used for biomarker discovery in this study. Paired tumor and nontumor TIF samples from 6 HBV-HCC patients were analyzed by a proteomic technique named iTRAQ (isobaric tag for relative and absolute quantitation). Totally, 241 up-regulated proteins (ratio ≥ 1.3, p < 0.05) and 288 down-regulated proteins (ratio ≤ −1.3, p < 0.05) in tumor TIF were identified. Interestingly, proteins in S100 family were found remarkably up-regulated in tumor TIF. One dramatically up-regulated protein S100A9 (ratio = 19) was further validated by ELISA in sera from liver cirrhosis (LC, HCC high risk population) and HCC patients (n = 47 for each group). The level of this protein was significantly elevated in HCC sera compared with LC (p < 0.0001). The area under the curve of this protein to distinguish HCC from LC was 0.83, with sensitivity of 91% (higher than AFP) and specificity of 66%. This result demonstrated the potential of S100A9 as a candidate HCC diagnostic biomarker. And TIF was a kind of promising material to identify candidate tumor biomarkers that could be detected in serum.

Serum soluble VSIG4 as a surrogate marker for the diagnosis of lymphoma‐associated hemophagocytic lymphohistiocytosis

“…Eight‐plex isobaric tags for relative and absolute quantitation (iTRAQ) analysis was performed in the State Key Laboratory of Proteomics (Beijing Proteome Research Center, National Center for Protein Sciences Beijing). Sample preparation, mass spectrometry, peptide and protein identification were performed as previously described (Wang et al ., ; Sun et al ., ; Zhang et al ., ). All reported data were recorded using Excel for evaluation of candidate proteins based on proteomic fold‐change and false discovery rate (FDR).…”

“…Usually, haemophagocytic macrophages in the bone marrow, peripheral blood or tissue samples are regarded as the hallmark of HLH, but these may not be present or be easily found at the early stage, leading to diagnostic difficulty (Zhang et al, 2011;Larroche, 2012;Maruoka et al, 2014). Particularly in the early stages of L-HLH, the clinical symptoms and laboratory results often do not formally fulfil the diagnostic HLH-2004 criteria, further leading to under-or delayed diagnosis of HLH in patients (Tamamyan et al, 2016;Lin et al, 2017).…”

“…All of their specimens were plasma samples, stored at À80°C and mailed to our laboratory on dry ice. performed as previously described Sun et al, 2016;Zhang et al, 2017). All reported data were recorded using Excel for evaluation of candidate proteins based on proteomic fold-change and false discovery rate (FDR).…”

“…Overall, 89 upregulated and 37 downregulated proteins were identified in L-HLH group compared with the non-HLH group (Table II). Among these soluble proteins upregulated in the serum, CD14, CD163, b2-microglobulin, and C-reactive protein (CRP) had been revealed to be present at higher levels in patients with HLH in previous studies (Bleesing et al, 2007;Nanno et al, 2016;Tamamyan et al, 2016;Koh et al, 2017;Machowicz et al, 2017). In addition to CD14 and CD163, other transmembrane proteins expressed on the macrophage's surface were also shown to be significantly upregulated in the serum of L-HLH, such as Fc gamma receptor III-A (FccRIIIA), scavenger receptor MARCO, CSF1 receptor (CSF1R) and VSIG4.…”

Discovering potential serological biomarker for chronic Hepatitis B Virus-related hepatocellular carcinoma in Chinese population by MAL-associated serum glycoproteomics analysis

“…demonstrated that versican core protein (VCAN) is a potential biomarker for early HCC diagnosis by label-free discovery analysis: selected (multiple) reaction monitoring (SRM/MRM)20. It was also reported that protein S100A9, which was dramatically up-regulated in sera from HCC patients, is a candidate HCC diagnostic biomarker21. Lu used the mass spectroscopic approach to characterize the metabolic features of the liver in HCC patients and verified that serum acetylcarnitine was a meaningful biomarker reflecting HCC diagnosis and progression22.…”