Screening and identification of apolipoprotein A-I as a potential hepatoblastoma biomarker in children, excluding inflammatory factors

Zhao,Wei; Li,Juan; Zhang,Yilin; Gao,Pengfei; Zhang,Junjie; Guo,Fei; Yu,Jiekai; Zheng,Shu; Wang,Jiaxiang

doi:10.3892/ol.2015.3207

Screening and identification of apolipoprotein A-I as a potential hepatoblastoma biomarker in children, excluding inflammatory factors

Authors:
- Wei Zhao
- Juan Li
- Yilin Zhang
- Pengfei Gao
- Junjie Zhang
- Fei Guo
- Jiekai Yu
- Shu Zheng
- Jiaxiang Wang
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Affiliations: Department of Pediatric Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Department of Respiratory Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China, Department of Clinical Medicine, Basic Medical College of Zhengzhou University, Zhengzhou, Henan 450001, P.R. China, Institute of Cancer, The Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China
Published online on: May 14, 2015 https://doi.org/10.3892/ol.2015.3207
Pages: 233-239
Copyright: © Zhao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to identify a child hepatoblastoma serum biomarker that is unaffected by inflammatory factors, with the ultimate aim of finding an effective method for the early diagnosis of hepatoblastoma. The magnetic bead‑based weak cation exchange chromatography technique was used to process serum harvested from 30 children with hepatoblastoma, 20 children with systemic inflammatory response syndrome (SIRS) and 20 healthy children. Proteins differentially expressed in SIRS were excluded from consideration as biomarkers for hepatoblastoma. Proteins differentially expressed in hepatoblastoma and healthy controls were screened using surface‑enhanced laser desorption/ionization‑time of flight‑mass spectrometry (SELDI‑TOF‑MS). Target proteins were purified by SDS-PAGE, and matrix‑assisted laser desorption/ionization (MALDI)‑TOF‑MS was used to determine their amino acid sequences. Protein matches were searched in the SwissProt database. Quantitative polymerase chain reaction (qPCR) and ELISA were employed to confirm the expression of target proteins. Following screening to exclude inflammatory factors, SELDI‑TOF‑MS revealed a protein with a mass‑to‑charge ratio of 9,348 Da that was expressed at significantly lower levels in the serum of children with hepatoblastoma compared with healthy controls (P<0.01). Sequence analysis identified this protein as apolipoprotein A-1 (Apo A‑I). qPCR and ELISA confirmed that the expression of Apo A‑I mRNA and protein were significantly lower in children with hepatoblastoma compared with healthy controls (P<0.05). These results indicate that Apo A‑I is a non‑inflammatory protein marker for hepatoblastoma with the potential for use in early diagnosis of hepatoblastoma. In addition, the present study demonstrates the feasibility of proteomic screening for the identification of proteins that can serve as markers for a specific tumor.

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