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BMC Cancer
Published in: BMC Cancer 1/2019

Open Access 01-12-2019 | Metastasis | Research article

Prognostic value of biomarkers EpCAM and αB-crystallin associated with lymphatic metastasis in breast cancer by iTRAQ analysis

Authors: Liang Zeng, Xiyun Deng, Jingmin Zhong, Li Yuan, Xiaojun Tao, Sai Zhang, Yong Zeng, Guangchun He, Pingping Tan, Yongguang Tao

Published in: BMC Cancer | Issue 1/2019

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Abstract

Background

Metastasis is responsible for the majority of deaths in a variety of cancer types, including breast cancer. Although several factors or biomarkers have been identified to predict the outcome of patients with breast cancer, few studies have been conducted to identify metastasis-associated biomarkers.

Methods

Quantitative iTRAQ proteomics analysis was used to detect differentially expressed proteins between lymph node metastases and their paired primary tumor tissues from 23 patients with metastatic breast cancer. Immunohistochemistry was performed to validate the expression of two upregulated (EpCAM, FADD) and two downregulated (NDRG1, αB-crystallin) proteins in 190 paraffin-embedded tissue samples. These four proteins were further analyzed for their correlation with clinicopathological features in 190 breast cancer patients.

Results

We identified 637 differentially regulated proteins (397 upregulated and 240 downregulated) in lymph node metastases compared with their paired primary tumor tissues. Data are available via ProteomeXchange with identifier PXD013931. Furthermore, bioinformatics analysis using GEO profiling confirmed the difference in the expression of EpCAM between metastases and primary tumors tissues. Two upregulated (EpCAM, FADD) and two downregulated (NDRG1, αB-crystallin) proteins were associated with the progression of breast cancer. Obviously, EpCAM plays a role in the metastasis of breast cancer cells to the lymph node. We further identified αB-crystallin as an independent biomarker to predict lymph node metastasis and the outcome of breast cancer patients.

Conclusion

We have identified that EpCAM plays a role in the metastasis of breast cancer cells to the lymph node. αB-crystallin, a stress-related protein that has recently been shown to be important for cell invasion and survival, was identified as a potential prognostic biomarker to predict the outcome of breast cancer patients.
Appendix
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Literature
1.
Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65(2):87–108.PubMed
2.
Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ, He J. Cancer statistics in China, 2015. CA Cancer J Clin. 2016;66(2):115–32.CrossRefPubMed
3.
Chen W, Zheng R, Zuo T, Zeng H, Zhang S, He J. National cancer incidence and mortality in China, 2012. Chin J Cancer Res. 2016;28(1):1–11.PubMedPubMedCentral
4.
Jin X, Mu P. Targeting breast Cancer metastasis. Breast Cancer (Auckl). 2015;9(Suppl 1):23–34.
5.
6.
DeMichele A, Yee D, Esserman L. Mechanisms of resistance to neoadjuvant chemotherapy in breast Cancer. N Engl J Med. 2017;377(23):2287–9.PubMedCrossRef
7.
Harper KL, Sosa MS, Entenberg D, Hosseini H, Cheung JF, Nobre R, Avivar-Valderas A, Nagi C, Girnius N, Davis RJ, et al. Mechanism of early dissemination and metastasis in Her2(+) mammary cancer. Nature. 2016;540(7634):588.PubMedPubMedCentralCrossRef
8.
Hosseini H, Obradovic MM, Hoffmann M, Harper KL, Sosa MS, Werner-Klein M, Nanduri LK, Werno C, Ehrl C, Maneck M, et al. Early dissemination seeds metastasis in breast cancer. Nature. 2016;540(7634):552.PubMedPubMedCentralCrossRef
9.
O'Shaughnessy J. Extending survival with chemotherapy in metastatic breast cancer. Oncologist. 2005;10(Suppl 3):20–9.PubMedCrossRef
10.
Spolverato G, Vitale A, Bagante F, Connolly R, Pawlik TM. Liver resection for breast Cancer liver metastases: a cost-utility analysis. Ann Surg. 2017;265(4):792–9.PubMedCrossRef
11.
Ohsfeldt RL, Ward MM, Schneider JE, Jaana M, Miller TR, Lei Y, Wakefield DS. Implementation of hospital computerized physician order entry systems in a rural state: feasibility and financial impact. J Am Med Inform Assoc. 2005;12(1):20–7.PubMedPubMedCentralCrossRef
12.
van de Vijver MJ, He YD, van't Veer LJ, Dai H, Hart AA, Voskuil DW, Schreiber GJ, Peterse JL, Roberts C, Marton MJ, et al. A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med. 2002;347(25):1999–2009.PubMedCrossRef
13.
Chandran UR, Ma C, Dhir R, Bisceglia M, Lyons-Weiler M, Liang W, Michalopoulos G, Becich M, Monzon FA. Gene expression profiles of prostate cancer reveal involvement of multiple molecular pathways in the metastatic process. BMC Cancer. 2007;7:64.PubMedPubMedCentralCrossRef
14.
Appierto V, Di Cosimo S, Reduzzi C, Pala V, Cappelletti V, Daidone MG. How to study and overcome tumor heterogeneity with circulating biomarkers: the breast cancer case. Semin Cancer Biol. 2017;44:106–16.PubMedCrossRef
15.
Ramaswamy S, Ross KN, Lander ES, Golub TR. A molecular signature of metastasis in primary solid tumors. Nat Genet. 2003;33(1):49–54.PubMedCrossRef
16.
Lorusso G, Ruegg C. New insights into the mechanisms of organ-specific breast cancer metastasis. Semin Cancer Biol. 2012;22(3):226–33.PubMedCrossRef
17.
Fry SA, Sinclair J, Timms JF, Leathem AJ, Dwek MV. A targeted glycoproteomic approach identifies cadherin-5 as a novel biomarker of metastatic breast cancer. Cancer Lett. 2013;328(2):335–44.PubMedCrossRef
18.
19.
Castle J, Shaker H, Morris K, Tugwood JD, Kirwan CC. The significance of circulating tumour cells in breast cancer: a review. Breast. 2014;23(5):552–60.PubMedCrossRef
20.
Lu X, Kang Y. Organotropism of breast cancer metastasis. J Mammary Gland Biol Neoplasia. 2007;12(2–3):153–62.PubMedCrossRef
21.
Patsialou A, Wang Y, Lin J, Whitney K, Goswami S, Kenny PA, Condeelis JS. Selective gene-expression profiling of migratory tumor cells in vivo predicts clinical outcome in breast cancer patients. Breast Cancer Res. 2012;14(5):R139.PubMedPubMedCentralCrossRef
22.
van’t Veer LJ, Dai H, van de Vijver MJ, He YD, Hart AA, Mao M, Peterse HL, van der Kooy K, Marton MJ, Witteveen AT, et al. Gene expression profiling predicts clinical outcome of breast cancer. Nature. 2002;415(6871):530–6.CrossRef
23.
Zieske LR. A perspective on the use of iTRAQ reagent technology for protein complex and profiling studies. J Exp Bot. 2006;57(7):1501–8.PubMedCrossRef
24.
Evans C, Noirel J, Ow SY, Salim M, Pereira-Medrano AG, Couto N, Pandhal J, Smith D, Pham TK, Karunakaran E, et al. An insight into iTRAQ: where do we stand now? Anal Bioanal Chem. 2012;404(4):1011–27.PubMedCrossRef
25.
Putz SM, Boehm AM, Stiewe T, Sickmann A. iTRAQ analysis of a cell culture model for malignant transformation, including comparison with 2D-PAGE and SILAC. J Proteome Res. 2012;11(4):2140–53.PubMedCrossRef
26.
Zeidan B, Manousopoulou A, Garay-Baquero DJ, White CH, Larkin SET, Potter KN, Roumeliotis TI, Papachristou EK, Copson E, Cutress RI, et al. Increased circulating resistin levels in early-onset breast cancer patients of normal body mass index correlate with lymph node negative involvement and longer disease free survival: a multi-center POSH cohort serum proteomics study. Breast Cancer Res. 2018;20(1):19.PubMedPubMedCentralCrossRef
27.
Bouchal P, Dvorakova M, Roumeliotis T, Bortlicek Z, Ihnatova I, Prochazkova I, Ho JT, Maryas J, Imrichova H, Budinska E, et al. Combined proteomics and transcriptomics identifies carboxypeptidase B1 and nuclear factor kappaB (NF-kappaB) associated proteins as putative biomarkers of metastasis in low grade breast Cancer. Mol Cell Proteomics. 2015;14(7):1814–30.PubMedPubMedCentralCrossRef
28.
Bouchal P, Roumeliotis T, Hrstka R, Nenutil R, Vojtesek B, Garbis SD. Biomarker discovery in low-grade breast cancer using isobaric stable isotope tags and two-dimensional liquid chromatography-tandem mass spectrometry (iTRAQ-2DLC-MS/MS) based quantitative proteomic analysis. J Proteome Res. 2009;8(1):362–73.PubMedCrossRef
29.
Jesneck JL, Mukherjee S, Yurkovetsky Z, Clyde M, Marks JR, Lokshin AE, Lo JY. Do serum biomarkers really measure breast cancer? BMC Cancer. 2009;9:164.PubMedPubMedCentralCrossRef
30.
Ruppen I, Grau L, Orenes-Pinero E, Ashman K, Gil M, Algaba F, Bellmunt J, Sanchez-Carbayo M. Differential protein expression profiling by iTRAQ-two-dimensional LC-MS/MS of human bladder cancer EJ138 cells transfected with the metastasis suppressor KiSS-1 gene. Mol Cell Proteomics. 2010;9(10):2276–91.PubMedPubMedCentralCrossRef
31.
Perez-Riverol Y, Csordas A, Bai J, Bernal-Llinares M, Hewapathirana S, Kundu DJ, Inuganti A, Griss J, Mayer G, Eisenacher M, et al. The PRIDE database and related tools and resources in 2019: improving support for quantification data. Nucleic Acids Res. 2019;47(D1):D442–50.PubMedCrossRef
32.
Ni J, Cozzi PJ, Duan W, Shigdar S, Graham PH, John KH, Li Y. Role of the EpCAM (CD326) in prostate cancer metastasis and progression. Cancer Metastasis Rev. 2012;31(3–4):779–91.PubMedCrossRef
33.
Pattje WJ, Melchers LJ, Slagter-Menkema L, Mastik MF, Schrijvers ML, Gibcus JH, Kluin PM, Hoegen-Chouvalova O, van der Laan BF, Roodenburg JL, et al. FADD expression is associated with regional and distant metastasis in squamous cell carcinoma of the head and neck. Histopathology. 2013;63(2):263–70.PubMedCrossRef
34.
Chang X, Xu X, Ma J, Xue X, Li Z, Deng P, Zhang S, Zhi Y, Chen J, Dai D. NDRG1 expression is related to the progression and prognosis of gastric cancer patients through modulating proliferation, invasion and cell cycle of gastric cancer cells. Mol Biol Rep. 2014;41(9):6215–23.PubMedCrossRef
35.
Liu S, Yan B, Lai W, Chen L, Xiao D, Xi S, Jiang Y, Dong X, An J, Chen X, et al. As a novel p53 direct target, bidirectional gene HspB2/alphaB-crystallin regulates the ROS level and Warburg effect. Biochim Biophys Acta. 2014;1839(7):592–603.PubMedCrossRef
36.
Cvekl A, McGreal R, Liu W. Lens development and Crystallin gene expression. Prog Mol Biol Transl Sci. 2015;134:129–67.PubMedCrossRef
37.
Haslbeck M, Peschek J, Buchner J, Weinkauf S. Structure and function of alpha-crystallins: Traversing from in vitro to in vivo. Biochim Biophys Acta. 2016;1860(1 Pt B):149–66.PubMedCrossRef
38.
Malin D, Strekalova E, Petrovic V, Deal AM, Al Ahmad A, Adamo B, Miller CR, Ugolkov A, Livasy C, Fritchie K, et al. alphaB-crystallin: a novel regulator of breast cancer metastasis to the brain. Clin Cancer Res. 2014;20(1):56–67.PubMedCrossRef
39.
Malin D, Strekalova E, Petrovic V, Rajanala H, Sharma B, Ugolkov A, Gradishar WJ, Cryns VL. ERK-regulated alphaB-crystallin induction by matrix detachment inhibits anoikis and promotes lung metastasis in vivo. Oncogene. 2015;34(45):5626–34.PubMedPubMedCentralCrossRef
40.
Petrovic V, Malin D, Cryns VL. alphaB-crystallin promotes oncogenic transformation and inhibits caspase activation in cells primed for apoptosis by Rb inactivation. Breast Cancer Res Treat. 2013;138(2):415–25.PubMedPubMedCentralCrossRef
41.
Wang F, Chen X, Li C, Sun Q, Chen Y, Wang Y, Peng H, Liu Z, Chen R, Liu K, et al. Pivotal role of augmented alphaB-crystallin in tumor development induced by deficient TSC1/2 complex. Oncogene. 2014;33(34):4352–8.PubMedCrossRef
42.
Chen Z, Ruan Q, Han S, Xi L, Jiang W, Jiang H, Ostrov DA, Cai J. Discovery of structure-based small molecular inhibitor of alphaB-crystallin against basal-like/triple-negative breast cancer development in vitro and in vivo. Breast Cancer Res Treat. 2014;145(1):45–59.PubMedCrossRef
43.
Huang Z, Cheng Y, Chiu PM, Cheung FM, Nicholls JM, Kwong DL, Lee AW, Zabarovsky ER, Stanbridge EJ, Lung HL, et al. Tumor suppressor alpha B-crystallin (CRYAB) associates with the cadherin/catenin adherens junction and impairs NPC progression-associated properties. Oncogene. 2012;31(32):3709–20.PubMedCrossRef
44.
Voduc KD, Nielsen TO, Perou CM, Harrell JC, Fan C, Kennecke H, Minn AJ, Cryns VL, Cheang MCU. alphaB-crystallin expression in breast Cancer is associated with brain metastasis. NPJ Breast Cancer. 2015;1:15014.PubMedPubMedCentralCrossRef
45.
Bandyopadhyay S, Pai SK, Hirota S, Hosobe S, Takano Y, Saito K, Piquemal D, Commes T, Watabe M, Gross SC, et al. Role of the putative tumor metastasis suppressor gene Drg-1 in breast cancer progression. Oncogene. 2004;23(33):5675–81.PubMedCrossRef
46.
Weigelt B, Peterse JL, van’t Veer LJ. Breast cancer metastasis: markers and models. Nat Rev Cancer. 2005;5(8):591–602.PubMedCrossRef
47.
Cimino Y, Costes A, Damotte D, Validire P, Mistou S, Cagnard N, Alifano M, Regnard JF, Chiocchia G, Sautes-Fridman C, et al. FADD protein release mirrors the development and aggressiveness of human non-small cell lung cancer. Br J Cancer. 2012;106(12):1989–96.PubMedPubMedCentralCrossRef
48.
Yoo NJ, Lee SH, Jeong EG, Lee JW, Soung YH, Nam SW, Kim SH, Lee JY, Lee SH. Expression of nuclear and cytoplasmic phosphorylated FADD in gastric cancers. Pathol Res Pract. 2007;203(2):73–8.PubMedCrossRef
49.
Tu W, Luo M, Wang Z, Yan W, Xia Y, Deng H, He J, Han P, Tian D. Upregulation of SATB1 promotes tumor growth and metastasis in liver cancer. Liver Int. 2012;32(7):1064–78.PubMedCrossRef
50.
Choi EJ, Yun JA, Jabeen S, Jeon EK, Won HS, Ko YH, Kim SY. Prognostic significance of TMEM16A, PPFIA1, and FADD expression in invasive ductal carcinoma of the breast. World J Surg Oncol. 2014;12:137.PubMedPubMedCentralCrossRef
51.
Bandyopadhyay S, Pai SK, Gross SC, Hirota S, Hosobe S, Miura K, Saito K, Commes T, Hayashi S, Watabe M, et al. The Drg-1 gene suppresses tumor metastasis in prostate cancer. Cancer Res. 2003;63(8):1731–6.PubMed
52.
Bandyopadhyay S, Wang Y, Zhan R, Pai SK, Watabe M, Iiizumi M, Furuta E, Mohinta S, Liu W, Hirota S, et al. The tumor metastasis suppressor gene Drg-1 down-regulates the expression of activating transcription factor 3 in prostate cancer. Cancer Res. 2006;66(24):11983–90.PubMedCrossRef
53.
Maruyama Y, Ono M, Kawahara A, Yokoyama T, Basaki Y, Kage M, Aoyagi S, Kinoshita H, Kuwano M. Tumor growth suppression in pancreatic cancer by a putative metastasis suppressor gene Cap43/NDRG1/Drg-1 through modulation of angiogenesis. Cancer Res. 2006;66(12):6233–42.PubMedCrossRef
54.
Cheng J, Xie HY, Xu X, Wu J, Wei X, Su R, Zhang W, Lv Z, Zheng S, Zhou L. NDRG1 as a biomarker for metastasis, recurrence and of poor prognosis in hepatocellular carcinoma. Cancer Lett. 2011;310(1):35–45.PubMedCrossRef
55.
Chang JT, Wang HM, Chang KW, Chen WH, Wen MC, Hsu YM, Yung BY, Chen IH, Liao CT, Hsieh LL, et al. Identification of differentially expressed genes in oral squamous cell carcinoma (OSCC): overexpression of NPM, CDK1 and NDRG1 and underexpression of CHES1. Int J Cancer. 2005;114(6):942–9.PubMedCrossRef
56.
Grover PK, Cummins AG, Price TJ, Roberts-Thomson IC, Hardingham JE. Circulating tumour cells: the evolving concept and the inadequacy of their enrichment by EpCAM-based methodology for basic and clinical cancer research. Ann Oncol. 2014;25(8):1506–16.PubMedCrossRef
57.
Gastl G, Spizzo G, Obrist P, Dunser M, Mikuz G. Ep-CAM overexpression in breast cancer as a predictor of survival. Lancet. 2000;356(9246):1981–2.PubMedCrossRef
58.
Ni J, Cozzi P, Hao J, Beretov J, Chang L, Duan W, Shigdar S, Delprado W, Graham P, Bucci J, et al. Epithelial cell adhesion molecule (EpCAM) is associated with prostate cancer metastasis and chemo/radioresistance via the PI3K/Akt/mTOR signaling pathway. Int J Biochem Cell Biol. 2013;45(12):2736–48.PubMedCrossRef
59.
Driemel C, Kremling H, Schumacher S, Will D, Wolters J, Lindenlauf N, Mack B, Baldus SA, Hoya V, Pietsch JM, et al. Context-dependent adaption of EpCAM expression in early systemic esophageal cancer. Oncogene. 2014;33(41):4904–15.PubMedCrossRef
60.
Romeu C, Farre X, Cardesa A, Nadal A. Expression of ep-CAM, but not of E48, associates with nodal involvement in advanced squamous cell carcinomas of the larynx. Histopathology. 2013;62(6):954–61.PubMedCrossRef
Metadata
Title
Prognostic value of biomarkers EpCAM and αB-crystallin associated with lymphatic metastasis in breast cancer by iTRAQ analysis
Authors
Liang Zeng
Xiyun Deng
Jingmin Zhong
Li Yuan
Xiaojun Tao
Sai Zhang
Yong Zeng
Guangchun He
Pingping Tan
Yongguang Tao
Publication date
01-12-2019
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2019
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-019-6016-3

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