Evaluation of HAAH/humbug quantitative detection in the diagnosis of hepatocellular carcinoma

Xue,Tian; Su,Jing; Li,Hongmin; Xue,Xiaoping

doi:10.3892/or.2014.3606

Evaluation of HAAH/humbug quantitative detection in the diagnosis of hepatocellular carcinoma

Authors:
- Tian Xue
- Jing Su
- Hongmin Li
- Xiaoping Xue
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Affiliations: Faculty of Life Science, Northwest University, Xi'an, Shaanxi 710069, P.R. China, Department of Microbiological Test, Nantong Center for Disease Control and Prevention, Nantong, Jiangsu 226007, P.R. China, Department of Microbiology, The Fourth Military Medical University, Xi'an, Shaanxi 710072, P.R. China
Published online on: November 13, 2014 https://doi.org/10.3892/or.2014.3606
Pages: 329-337

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Abstract

Human aspartyl-(asparaginyl)-β-hydroxylase (HAAH) is a type 2 transmembrane protein and an α-ketoglutarate-dependent dioxygenase that can stereospecifically catalyze the post-translational hydroxylation reaction of β-carbon atoms of aspartic acid and asparagine residues present in epidermal growth factor-like domains of certain specific proteins. Humbug is a truncated isoform of aspartyl (asparaginyl) β-hydroxylase that lacks the catalytic domain. A series of reports demonstrated that overexpression of HAAH/humbug was identified in hepatocellular carcinoma (HCC) and various tumor tissues. However, the prognostic value of HAAH/humbug expression in HCC remains unclear. The purpose of this study was to investigate the expression of the HAAH/humbug gene at the mRNA and protein levels in HCC and to assess the overexpression of HAAH/humbug as a diagnostic and prognostic marker in HCC. HAAH/humbug mRNA levels were measured in 120 HCCs and 40 paired non-tumor liver tissues by molecular beacon (MB) quantitative RT-PCR. Immunohistochemical staining was used to detect the protein level of the HAAH/humbug in the same specimens. ROC analysis was performed based on the expression levels of the HAAH/humbug gene in 120 cases of HCC tissues and 40 cases of adjacent non-tumor liver tissues. The results showed that 117 (97.5%) of the 120 frozen sections of patients with HCCs had HAAH/humbug-positive immunoreactivity, whereas the 40 adjacent non-tumor liver tissues exhibited no staining. Higher levels of HAAH/humbug mRNA were found in 114 (95%) of the 120 HCC tissues relative to the adjacent cancer‑free tissue. ROC curve analysis exhibited that the sensitivity was 90.1%, specificity was 97.6% and ROC AUC was 0.986. The specific value of HAAH/β-actin abundance used as a cut‑off point was 0.315, while the gene copy number (7.35 copies/µl) was used a as cut‑off point, with sensitivity being 99.2%, specificity 96.7% and the ROC AUC used 0.990. No statistically significant difference was observed for these two factors. HAAH/humbug expression levels were upregulated in almost all the HCC tissues when compared to the adjacent cancer-free tissue, irrespective of the cut‑off point used. Results of the present study suggested that HAAH/humbug is a potential diagnostic and prognostic biomarker for the treatment of HCC.

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