Multiplexed Proteomic Approach for Identification of Serum Biomarkers in Hepatocellular Carcinoma Patients with Normal AFP

Abstract

Alpha fetoprotein (AFP) has been used as a serologic indicator of hepatocellular carcinoma (HCC). We aimed to identify an HCC-specific serum biomarker for diagnosis using a multiplexed proteomic technique in HCC patients with normal AFP levels. A total of 152 patients were included from Guro Hospital, Korea University. Among 267 identified proteins, 28 and 86 proteins showed at least a two-fold elevation or reduction in expression, respectively. Multiple reaction monitoring (MRM) analysis of 41 proteins revealed 10 proteins were differentially expressed in patients with liver cirrhosis and HCC patients with normal AFP. A combination of tripartite motif22 (Trim22), seprase, and bone morphogenetic protein1 had an area under receiver operating characteristic of 0.957 for HCC diagnosis. Real-time PCR and western blot analysis of the paired tumor/non-tumor liver tissue in HCC revealed a reduced expression of Trim22 in the tumor tissue. Also, serum levels of Trim22 were significantly reduced in HCC patients with normal AFP compared to those with liver cirrhosis (p = 0.032). Inhibition of Trim22 increased cellular proliferation in human hepatoma cell lines, whereas overexpression of Trim22 decreased cellular proliferation in hepatoma cell lines. In conclusion, the combination of three serum markers improved the chance of diagnosing HCC. MRM-based quantification of the serum protein in patients with normal AFP provides the potential for early diagnosis of HCC.

Advances in High Throughput Proteomics Profiling in Establishing Potential Biomarkers for Gastrointestinal Cancer

“…For example, Heo et al achieved specificity and sensitivity of around 80% for early diagnostic HCC marker EIF3A peptides from serum samples, and ELISA and western blotting validation confirmed the use of selected peptides clinically [100]. Similar to the previous study performed by Zhan's group [99], Lee's group also validated the presence of AFP in a large number of HCC patients' serum samples and verified AFP expression by using RT-qPCR and western blotting techniques [101]. Furthermore, another study revealed that AFP along with heptoglobin specific N-glycopeptides may achieve a sensitivity of greater than 80% as diagnostic biomarkers for nonalcoholic steatohepatitis HCC patients [102].…”

Proteome Multimarker Panel for the Early Detection of Hepatocellular Carcinoma: Multicenter Derivation, Validation, and Comparison

“…No study that has aimed to identify new HCC serum markers through mass spectrometry − has validated the discriminatory performance of the identified markers in a chronologically separately collected, multicenter cohort. The current study is also distinct from a previous report by our group, in that data preprocessing was adopted before the selection of the markers and that fewer markers were involved in the developed panel.…”

Clinical Value and Underlying Mechanisms of Upregulated LINC00485 in Hepatocellular Carcinoma

“…Nowadays, Ultrasonography and AFP detection are commonly accepted as useful methods to periodically screen HCC (17)(18)(19). Imaging diagnosis of early HCC still remains some troubles, and serum AFP has a nonnegligible false negative rate of about 30%, which could potentially lead to misdiagnosis, missed diagnosis, and a delay in timely treatment (20)(21)(22). Therefore, we planned to analyze whether LINC00485 can help clinicians better distinguish HCC patients.…”