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Archive > Volume 32 Issue 3 > 2025,32(3):270-280. DOI:10.3872/j.issn.1007-385X.2025.03.006 Prev Next
Prompt Report
The expression of early hepatocellular carcinoma-related antigen CTAG1A in hepatocellular carcinoma tissues and cells and identification of cytotoxic T lymphocyte epitopes
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Abstract:
[Abstract] Objective: Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver. The diagnosis rate of early HCC is low, and most patients are diagnosed at the late stage and have a very poor prognosis. Therefore, it is urgent to explore effective early diagnosis markers and intervention targets for HCC. Cancer/testicular antigen 1A (CTAG1A) is abnormally expressed and highly immunogenic in a variety of tumors, but its expression characteristics and immunogenicity in HCC remain unclear. The aim of this study is to identify the expression and immunogenicity of CTAG1A in HCC tissues and cells, providing a new biomarker for the early diagnosis of HCC and a new potential target for clinical immunotherapy. Methods: This study screened the differentially expressed gene profiles between 10 pairs of very early HCC (BCLC stage 0 HCC) tumors and paracancerous tissues using a transcriptome microarray. The expression of CTAG1A was verified by RT-qPCR in an independent large sample (BCLC stage 0, A, B, C HCC tissues and adjacent non-tumor tissues, n=149) and various hepatocellular carcinoma cell lines. Bioinformatics tools (TepiTool of IEDB database and Swiss Model) were used to predict the MHC-Ⅰ and MHC-Ⅱ epitopes of CTAG1A. The candidate peptides were synthesized by solid-phase polypeptide synthesis method. After purification by HPLC and verification by mass spectrometry assay, the specific T cell responses of peripheral blood mononuclear cells (PBMC) of 9 HCC patients to all peptides were detected by IFN-γ enzyme-linked immunospot assay (ELISpot). The clinical samples were collected from HCC patients admitted to the Third Affiliated Hospital of Naval Medical University (Eastern Hepatobiliary Surgery Hospital) from 2015 to 2022. The collection and usage of all samples were carried out with the consent of the patients, and with the approval of the Ethics Committee of Eastern Hepatobiliary Surgery Hospital (EHBHKY2015-01-017) and in strict accordance with relevant requirements and ethical regulations. Statistical analysis was performed using SPSS 30.0 software, and the diagnostic efficiency was evaluated by ROC curve. Results: Transcriptome chip screening results showed that CTAG1A expression was significantly up-regulated in the very early-stage HCC (BCLC stage 0 HCC) (|FC| = 99.16, P < 0.0001). The verification using the clinical independent samples showed its high expression in all stages of HCC and better diagnostic efficacy in early-stage HCC (BCLC stage 0 HCC AUC=0.6893, sensitivity = 85.71%; BCLC stage A HCC AUC = 0.8229, sensitivity = 83.33%). Furthermore, the expression of CTAG1A was significantly higher in multiple liver cancer cell lines than in relatively normal liver cell lines (P < 0.001). Compared with alpha-fetoprotein (AFP), CTAG1A showed better diagnostic efficacy in BCLC stage 0 and stage A HCC (ROC curve analysis of AFP showed no significant difference in early HCC, P > 0.05). Bioinformatics tools predicted that CTAG1A contained 8 MHC-type I and 4 MHC-type II epitopes. The IFN-γ ELISpot assay showed that 12 synthetic peptides could induce PBMC specific T cell response in HCC patients to varying degrees. Conclusion: CTAG1A is significantly overexpressed in early-stage HCC and has multi-epitope immunogenicity, which may activate CD8? and CD4? T cells, suggesting its potential as a target for HCC immunotherapy. It may provide a new direction for developing combined immunotherapy strategies based on mRNA vaccines or adoptive cell therapy. Compared with AFP, CTAG1A exhibits better diagnostic efficacy in early stage HCC, suggesting its potential as a marker for early diagnosis of HCC.
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