Serum and tissue proteomic signatures of patients with hepatocellular carcinoma using 2‑D gel electrophoresis

Peng,Huifang; Yan,Zhijian; Zeng,Xinhua; Zhang,Sheng; Jiang,Hongwei; Huang,Heqing; Zhuo,Huiqin

doi:10.3892/mmr.2019.10311

Serum and tissue proteomic signatures of patients with hepatocellular carcinoma using 2‑D gel electrophoresis

Authors:
- Huifang Peng
- Zhijian Yan
- Xinhua Zeng
- Sheng Zhang
- Hongwei Jiang
- Heqing Huang
- Huiqin Zhuo
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Affiliations: Department of Endocrinology, The First Affiliated Hospital and College of Clinical Medicine of Henan University of Science and Technology, Luoyang, Henan 471000, P.R. China, Department of Urology, The Affiliated Zhongshan Hospital, Xiamen University, Xiamen, Fujian 361004, P.R. China, State Key Laboratory of Stress Cell Biology, School of Life Science, Xiamen University, Xiamen, Fujian 361004, P.R. China, Fujian Provincial Key Laboratory of Chronic Liver Disease and Hepatocellular Carcinoma, The Affiliated Zhongshan Hospital, Xiamen University, Xiamen, Fujian 361004, P.R. China, Department of Gastrointestinal Surgery, Zhongshan Hospital, Xiamen University, Xiamen, Fujian 361004, P.R. China
Published online on: May 29, 2019 https://doi.org/10.3892/mmr.2019.10311
Pages: 1025-1038
Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hepatocellular carcinoma (HCC) accounts for ~85% of primary liver cancer cases and is a leading cause of mortality worldwide. Effective early diagnosis is difficult for HCC; however, effective biomarkers may be beneficial for diagnosis. In the current study, serum samples, and HCC and adjacent tissue samples were obtained from patients with HCC for the detection of biomarkers using 2‑D gel electrophoresis (2‑DE) and matrix‑assisted laser desorption/ionization‑time of flight (TOF)/TOF mass spectrometry. The crude serum samples did not need to be prepared for removal of high abundance proteins. The mRNA expression levels of HCC‑associated proteins were detected in tissues using reverse transcription‑quantitative PCR. Statistical analysis and database matching were used to identify the differentially expressed proteins detected in the serum and tissue groups. Immunohistochemistry (IHC) was performed to detect the expression of significant proteins in HCC and adjacent tissues. The results revealed ~800 protein spots on a 2‑DE gel that were detected in serum samples, and 1,200 spots were identified in the tissue samples. The protein and mRNA expression levels of oxysterol binding protein‑like 11 (OSBPL11) in HCC serum and tissue samples were consistent. Pathway analysis demonstrated that members of the apolipoprotein family, particularly apolipoprotein E (APOE), and RAS family members were closely associated in HCC, either directly or via ferratin heavy polypeptide 1. IHC results demonstrated that the APOE protein serves an important role in liver cancer development. The lysis buffer used in the current study was effective for serum protein separation in 2‑DE sample preparation. In addition, the present study revealed that downregulated OSBPL11 may be a potential indicator for HCC, and the apolipoprotein family, particularly APOE, and the RAS family may cooperatively serve an important role.

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1	Li S, Yang F and Ren X: Immunotherapy for hepatocellular carcinoma. Drug Discov Ther. 9:363–371. 2015. View Article : Google Scholar : PubMed/NCBI
2	McGlynn KA, Petrick JL and London WT: Global epidemiology of hepatocellular carcinoma: An emphasis on demographic and regional variability. Clin Liver Dis. 19:223–238. 2015. View Article : Google Scholar : PubMed/NCBI
3	Torre LA, Bray F, Siegel R, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
4	Han L, Lv Y, Guo H, Ruan Z and Nan K: Implications of biomarkers in human hepatocellular carcinoma pathogenesis and therapy. World J Gastroenterol. 20:10249–10261. 2014. View Article : Google Scholar : PubMed/NCBI
5	Ray S, Patel SK, Kumar V, Damahe J and Srivastava S: Differential expression of serum/plasma proteins in various infectious diseases: Specific or nonspecific signatures. Proteomics Clin Appl. 8:53–72. 2014. View Article : Google Scholar : PubMed/NCBI
6	Chromy BA, Gonzales AD, Perkins J, Choi MW, Corzett MH, Chang BC, Corzett CH and McCutchen-Maloney SL: Proteomic analysis of human serum by two-dimensional differential gel electrophoresis after depletion of high-abundant proteins. J Proteome Res. 3:1120–1127. 2014. View Article : Google Scholar
7	Sano S, Tagami S, Hashimoto Y, Yoshizawa-Kumagaye K, Tsunemi M, Okochi M and Tomonaga T: Absolute quantitation of low abundance plasma APL1β peptides at sub-fmol/mL Level by SRM/MRM without immunoaffinity enrichment. J Proteome Res. 13:1012–1020. 2014. View Article : Google Scholar : PubMed/NCBI
8	Xu S, Liu P, Lu X, Zhang J, Huang L, Hua W, He D and Ouyang J: A highly sensitive ‘turn-on’ fluorescent sensor for the detection of human serum proteins based on the size exclusion of the polyacrylamide gel. Electrophoresis. 35:546–553. 2014. View Article : Google Scholar : PubMed/NCBI
9	Luo LZ, Jin HW and Huang HQ: Transferrin-cisplatin specifically deliver cisplatin to HepG2 cells in vitro and enhance cisplatin cytotoxicity. J Proteomics. 77:237–250. 2012. View Article : Google Scholar : PubMed/NCBI
10	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
11	Liu W, Liu B, Cai Q, Li J, Chen X and Zhu Z: Proteomic identification of serum biomarkers for gastric cancer using multi-dimensional liquid chromatography and 2D differential gel electrophoresis. Clin Chim Acta. 413:1098–1106. 2012. View Article : Google Scholar : PubMed/NCBI
12	Qin S, Ferdinand AS, Richie JP, O'Leary MP, Mok SC and Liu BC: Chromatofocusing fractionation and two-dimensional difference gel electrophoresis for low abundance serum proteins. Proteomics. 5:3183–3192. 2005. View Article : Google Scholar : PubMed/NCBI
13	Chen YY, Lin SY, Yeh YY, Hsiao HH, Wu CY, Chen ST and Wang AH: A modified protein precipitation procedure for efficient removal of albumin from serum. Electrophoresis. 26:2117–2127. 2005. View Article : Google Scholar : PubMed/NCBI
14	Amraoui F, Hassani Lahsinoui H, Boussata S, Keijser R, Veenboer GJ, Middeldorp S, van der Post JA, Ris-Stalpers C, Afink GB and van den Born BJ: Placental expression of heparan sulfate 3-O-sulfotransferase-3A1 in normotensive and pre-eclamptic pregnancies. Placenta. 36:1218–1224. 2015. View Article : Google Scholar : PubMed/NCBI
15	Gajewski MM, Alisaraie L and Tuszynski JA: Peloruside, laulimalide, and noscapine interactions with beta-tubulin. Pharm Res. 29:2985–2993. 2012. View Article : Google Scholar : PubMed/NCBI
16	Hall RE, Horsfall DJ, Stahl J, Vivekanandan S, Ricciardelli C, Stapleton AM, Scardino PT, Neufing P and Tilley WD: Apolipoprotein-D: A novel cellular marker for HGPIN and prostate cancer. Prostate. 58:103–108. 2004. View Article : Google Scholar : PubMed/NCBI
17	Lau KS and Haigis KM: Non-redundancy within the RAS oncogene family: Insights into mutational disparities in cancer. Mol Cells. 28:315–320. 2009. View Article : Google Scholar : PubMed/NCBI
18	Chang TW, Chen CC, Chen KY, Su JH, Chang JH and Chang MC: Ribosomal phosphoprotein P0 interacts with GCIP and overexpression of P0 is associated with cellular proliferation in breast and liver carcinoma cells. Oncogene. 27:332–338. 2008. View Article : Google Scholar : PubMed/NCBI
19	Weber-Boyvat M, Zhong W, Yan D and Olkkonen VM: Oxysterol-binding proteins: Functions in cell regulation beyond lipid metabolism. Biochem Pharmacol. 86:89–95. 2013. View Article : Google Scholar : PubMed/NCBI
20	Olkkonen VM and Levine TP: Oxysterol binding proteins: In more than one place at one time? Biochem Cell Biol. 82:87–98. 2004. View Article : Google Scholar : PubMed/NCBI
21	Olkkonen VM: OSBP-related protein family in lipid transport over membrane contact sites. Lipid Insights. 8 (Suppl 1):S1–S9. 2015.
22	Olkkonen VM and Li S: Oxysterol-binding proteins: Sterol and phosphoinositide sensors coordinating transport, signaling and metabolism. Prog Lipid Res. 52:529–538. 2013. View Article : Google Scholar : PubMed/NCBI
23	Bondi J, Husdal A, Bukholm G, Nesland JM, Bakka A and Bukholm IR: Expression and gene amplification of primary (A, B1, D1, D3, and E) and secondary (C and H) cyclins in colon adenocarcinomas and correlation with patient outcome. J Clin Pathol. 58:509–514. 2005. View Article : Google Scholar : PubMed/NCBI
24	Caraglia M, Park MH, Wolff EC, Marra M and Abbruzzese A: eIF5A isoforms and cancer: Two brothers for two functions? Amino Acids. 44:103–109. 2013. View Article : Google Scholar : PubMed/NCBI
25	Mathews MB and Hershey JW: The translation factor eIF5A and human cancer. Biochim Biophys Acta. 1849:836–844. 2015. View Article : Google Scholar : PubMed/NCBI
26	Stiborová M, Indra R, Frei E, Kopečková K, Schmeiser HH, Eckschlager T, Adam V, Heger Z, Arlt VM and Martínek V: Cytochrome b5 plays a dual role in the reaction cycle of cytochrome P450 3A4 during oxidation of the anticancer drug ellipticine. Monatshefte Fur Chemie. 148:1983–1991. 2017. View Article : Google Scholar : PubMed/NCBI
27	Henderson CJ, McLaughlin LA, Finn RD, Ronseaux S, Kapelyukh Y and Wolf CR: A role for cytochrome b5 in the in vivo disposition of anticancer and cytochrome P450 probe drugs in mice. Drug Metab Dispos. 42:70–77. 2014. View Article : Google Scholar : PubMed/NCBI
28	Chekhun SV, Lukyanova NY, Shvets YV, Burlaka AP and Buchinska LG: Significance of ferritin expression in formation of malignant phenotype of human breast cancer cells. Exp Oncol. 36:179–183. 2014.PubMed/NCBI
29	Jiang XP and Elliott RL: Decreased iron in cancer cells and their microenvironment improves cytolysis of breast cancer cells by natural killer cells. Anticancer Res. 37:2297–2305. 2007. View Article : Google Scholar
30	Dai Z, Zhou SL, Zhou ZJ, Bai DS, Xu XY, Fu XT, Chen Q, Zhao YM, Zhu K, Yu L, et al: Capn4 contributes to tumour growth and metastasis of hepatocellular carcinoma by activation of the FAK-Src signalling pathways. J Pathol. 234:316–328. 2014. View Article : Google Scholar : PubMed/NCBI
31	Leal MF, Calcagno DQ, Demachki S, Assumpcao PP, Chammas R, Burbano RR and Smith Mde A: Clinical implication of 14-3-3 epsilon expression in gastric cancer. World J Gastroenterol. 18:1531–1537. 2012. View Article : Google Scholar : PubMed/NCBI
32	Yang Y, Cai J, Yin J, Wang D, Bai Z, Zhang J, Wang K, Yu G and Zhang Z: Inorganic pyrophosphatase (PPA1) is a negative prognostic marker for human gastric cancer. Int J Clin Exp Pathol. 8:12482–12490. 2015.PubMed/NCBI
33	Tang S, Huang W, Zhong M, Yin L, Jiang H, Hou S, Gan P and Yuan Y: Identification Keratin 1 as a cDDP-resistant protein in nasopharyngeal carcinoma cell lines. J Proteomics. 75:2352–2360. 2012. View Article : Google Scholar : PubMed/NCBI
34	Jing Z, Yin H, Wang P, Gao J and Yuan L: Centlein, a novel microtubule-associated protein stabilizing microtubules and involved in neurite formation. Biochem Biophys Res Commun. 472:360–365. 2016. View Article : Google Scholar : PubMed/NCBI
35	Su WP, Chen YT, Lai WW, Lin CC, Yan JJ and Su WC: Apolipoprotein E expression promotes lung adenocarcinoma proliferation and migration and as a potential survival marker in lung cancer. Lung Cancer. 71:28–33. 2011. View Article : Google Scholar : PubMed/NCBI
36	Hulpke S, Baldauf C and Tampe R: Molecular architecture of the MHC I peptide-loading complex: One tapasin molecule is essential and sufficient for antigen processing. FASEB J. 26:5071–5080. 2012. View Article : Google Scholar : PubMed/NCBI