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. 2020 Jan 23:2020:2632716.
doi: 10.1155/2020/2632716. eCollection 2020.

Identification of Protein Expression Changes in Hepatocellular Carcinoma through iTRAQ

Yuanyuan Zhang  1 , Xia Ying  2 , Qian Zhao  3 , Jinlu Ma  4 , Dan Zhang  5 , Chenchen He  4 , Suxia Han  4
Affiliations

Identification of Protein Expression Changes in Hepatocellular Carcinoma through iTRAQ

Yuanyuan Zhang et al. Dis Markers. .

Abstract

Background: Hepatocellular carcinoma (HCC) is a malignant tumor associated with a poor prognosis. Serum biomarkers of HCC have the potential to improve the diagnosis, provide a means to monitor the tumors, and predict their malignancy. Proteins that are expressed differentially between HCC patients and normal controls have the potential to be biomarkers.

Method: Serum samples from 10 confirmed HCC patients and 10 controls were collected. The differentially expressed proteins in the serum were identified using an isobaric tags for relative and absolute quantitation- (iTRAQ-) based method. Potential serum biomarkers were validated by ELISA in another 20 HCC patients and 20 controls. Their expression data in HCC were extracted from The Cancer Genome Atlas (TCGA) dataset.

Results: A total of 260 proteins were measured in the serum of HCC patients and compared to those in sex- and age-matched normal controls. Forty-one proteins displayed significant changes, with 26 being downregulated and 15 being upregulated. Upregulated proteins included alpha-1-antitrypsin (A1AT) and peroxiredoxin 2 (PRDX2), and downregulated proteins included paraoxonase 1 (PON1) and C-reactive protein (CRP). We then used ELISA to measure serum levels of A1AT, PRDX2, PON1, and CRP in another 20 patients with HCC and found that only PON1 levels were consistent with the iTRAQ result. In TCGA dataset, PON1 expression was downregulated in HCC tissues (P < 0.001) and low expression of PON1 was associated with poor survival in HCC patients (P < 0.001) and low expression of PON1 was associated with poor survival in HCC patients (.

Conclusions: PON1 could act as a biomarker for HCC to assist in the diagnosis of HCC.

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Conflict of interest statement

The authors who took part in this study declare that they do not have anything to disclose regarding funding or a conflict of interest with respect to this manuscript.

Figures

Figure 1
Figure 1
(a) Differentially expressed serum proteins in HCC patients detected by iTRAQ. (b) Cellular components of the differentially expressed proteins. (c) Biological processes involved by the differentially expressed proteins. (d) Molecular functions of the differentially expressed proteins. (e) KEGG pathway analysis of the differentially expressed proteins.
Figure 2
Figure 2
Validation of differentially expressed proteins in HCC serum by ELISA. Serum levels of (a) alpha-1-antitrypsin (A1AT), (b) peroxiredoxin 2 (PRDX2), (c) paraoxonase 1 (PON1), and (d) C-reactive protein (CRP) in HCC patients and normal controls. ∗∗P < 0.01 vs. control.
Figure 3
Figure 3
Hepatocellular carcinoma (HCC) data of (a) alpha-1-antitrypsin (A1AT), (b) peroxiredoxin 2 (PRDX2), (c) paraoxonase 1 (PON1), and (d) C-reactive protein (CRP) expression in The Cancer Genome Atlas (TCGA) database. Data are presented as medians (interquartile range (IQR)) and compared by the Mann-Whitney rank-sum test.
Figure 4
Figure 4
The relationship between the survival of HCC patients and the expression levels of (a) alpha-1-antitrypsin (A1AT), (b) peroxiredoxin 2 (PRDX2), (c) paraoxonase 1 (PON1), and (d) C-reactive protein (CRP) in TCGA database. The expression levels were divided into the high-expression and low-expression groups based on the maximally separated Kaplan-Meier plots.
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