Nuclear EpICD expression and its role in hepatocellular carcinoma

Park,Shin  Young; Bae,Jun  Sang; Cha,Eun  Jung; Chu,Hyun  Hee; Sohn,Jang  Sihn; Moon,Woo  Sung

doi:10.3892/or.2016.4789

Nuclear EpICD expression and its role in hepatocellular carcinoma

Authors:
- Shin Young Park
- Jun Sang Bae
- Eun Jung Cha
- Hyun Hee Chu
- Jang Sihn Sohn
- Woo Sung Moon
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Affiliations: Department of Pathology, Konyang University College of Medicine, Daejeon 35365, Republic of Korea, Department of Pathology, Chonbuk National University, Medical School Research Institute of Clinical Medicine of Chonbuk National University-Biomedical Research Institute of Chonbuk National University Hospital and Research Institute for Endocrine Sciences, Jeonju 561-756, Republic of Korea
Published online on: May 6, 2016 https://doi.org/10.3892/or.2016.4789
Pages: 197-204

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Abstract

Regulated intramembrane proteolysis of epithelial cell adhesion molecule (EpCAM) results in shedding of the extracellular domain (EpEX) and release of the intracellular domain (EpICD) into the cytoplasm. Released EpICD associates with FHL2, β-catenin and Lef-1 to form a nuclear complex and triggers oncogenic signaling. This study was conducted to examine the nuclear expression of EpICD in hepatocellular carcinoma (HCC) and to assess the role of EpICD in HCC. EpICD immunoexpression was examined in 100 cases of HCC using tissue microarrays and correlated with clinicopathological parameters. We also examined the role of EpICD in HCC using EpICD cDNA transfected HCC cell line and EpCAM silenced HCC cell line by small interfering RNA (siRNA). Nuclear expression of EpICD was observed in 19 of 100 (19%) cases. Nuclear expression of EpICD significantly correlated with nuclear expression of β-catenin, and Ki-67 labeling index. In addition, nuclear expression of EpICD was associated with higher histologic grade and advanced T category. Forced overexpression of EpICD in the HCC cell significantly increased the cell proliferation, migration and invasion. The overexpression of EpICD also increased the expression levels of the active form of β-catenin and c-myc and cyclin D1. In contrast, downregulation of EpCAM by siRNA decreased the cell proliferation, migration, invasion and the expression of active form of β-catenin, c-myc and cyclin D1. Our present data suggest that EpICD plays important roles in HCC progression by modulating expression of target genes of EpCAM.

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1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar
2	Graf D, Vallbohmer D, Knoefel WT, Kröpil P, Antoch G, Sagir A and Häussinger D: Multimodal treatment of hepatocellular carcinoma. Eur J Intern Med. 25:430–437. 2014. View Article : Google Scholar : PubMed/NCBI
3	Litvinov SV, Velders MP, Bakker HA, Fleuren GJ and Warnaar SO: Ep-CAM: A human epithelial antigen is a homophilic cell-cell adhesion molecule. J Cell Biol. 125:437–446. 1994. View Article : Google Scholar : PubMed/NCBI
4	Baeuerle PA and Gires O: EpCAM (CD326) finding its role in cancer. Br J Cancer. 96:417–423. 2007. View Article : Google Scholar : PubMed/NCBI
5	van der Gun BT, Melchers LJ, Ruiters MH, de Leij LF, McLaughlin PM and Rots MG: EpCAM in carcinogenesis: The good, the bad or the ugly. Carcinogenesis. 31:1913–1921. 2010. View Article : Google Scholar : PubMed/NCBI
6	Went PT, Lugli A, Meier S, Bundi M, Mirlacher M, Sauter G and Dirnhofer S: Frequent EpCam protein expression in human carcinomas. Hum Pathol. 35:122–128. 2004. View Article : Google Scholar : PubMed/NCBI
7	Schmidt M, Scheulen ME, Dittrich C, Obrist P, Marschner N, Dirix L, Schmidt M, Ruttinger D, Schuler M, Reinhardt C, et al: An open-label, randomized phase II study of adecatumumab, a fully human anti-EpCAM antibody, as monotherapy in patients with metastatic breast cancer. Ann Oncol. 21:275–282. 2010. View Article : Google Scholar
8	Niedzwiecki D, Bertagnolli MM, Warren RS, Compton CC, Kemeny NE, Benson AB, Eckhardt SG, Alberts S, Porjosh GN, Kerr DJ, et al: Documenting the natural history of patients with resected stage II adenocarcinoma of the colon after random assignment to adjuvant treatment with edrecolomab or observation: Results from CALGB 9581. J Clin Oncol. 29:3146–3152. 2011. View Article : Google Scholar : PubMed/NCBI
9	Maetzel D, Denzel S, Mack B, Canis M, Went P, Benk M, Kieu C, Papior P, Baeuerle PA, Munz M, et al: Nuclear signalling by tumour-associated antigen EpCAM. Nat Cell Biol. 11:162–171. 2009. View Article : Google Scholar : PubMed/NCBI
10	Ralhan R, Cao J, Lim T, Macmillan C, Freeman JL and Walfish PG: EpCAM nuclear localization identifies aggressive thyroid cancer and is a marker for poor prognosis. BMC Cancer. 10:3312010. View Article : Google Scholar : PubMed/NCBI
11	Ralhan R, He HC, So AK, Tripathi SC, Kumar M, Hasan MR, Kaur J, Kashat L, MacMillan C, Chauhan SS, et al: Nuclear and cytoplasmic accumulation of Ep-ICD is frequently detected in human epithelial cancers. PLoS One. 5:e141302010. View Article : Google Scholar : PubMed/NCBI
12	Jachin S, Bae JS, Sung JJ, Park HS, Jang KY, Chung MJ, Kim DG and Moon WS: The role of nuclear EpICD in extrahepatic cholangiocarcinoma: Association with β-catenin. Int J Oncol. 45:691–698. 2014.PubMed/NCBI
13	Srivastava G, Assi J, Kashat L, Matta A, Chang M, Walfish PG and Ralhan R: Nuclear Ep-ICD accumulation predicts aggressive clinical course in early stage breast cancer patients. BMC Cancer. 14:7262014. View Article : Google Scholar : PubMed/NCBI
14	Kim DG, Park SY, Kim H, Chun YH, Moon WS and Park SH: A comprehensive karyotypic analysis on a newly established sarcomatoid hepatocellular carcinoma cell line SH-J1 by comparative genomic hybridization and chromosome painting. Cancer Genet Cytogenet. 132:120–124. 2002. View Article : Google Scholar : PubMed/NCBI
15	Dollé L, Theise ND, Schmelzer E, Boulter L, Gires O and van Grunsven LA: EpCAM and the biology of hepatic stem/progenitor cells. Am J Physiol Gastrointest Liver Physiol. 308:G233–G250. 2015. View Article : Google Scholar :
16	Chan AW, Tong JH, Chan SL, Lai PB and To KF: Expression of stemness markers (CD133 and EpCAM) in prognostication of hepatocellular carcinoma. Histopathology. 64:935–950. 2014. View Article : Google Scholar : PubMed/NCBI
17	Winter MJ, Nagelkerken B, Mertens AE, Rees-Bakker HA, Briaire-de Bruijn IH and Litvinov SV: Expression of Ep-CAM shifts the state of cadherin-mediated adhesions from strong to weak. Exp Cell Res. 285:50–58. 2003. View Article : Google Scholar : PubMed/NCBI
18	Chaves-Pérez A, Mack B, Maetzel D, Kremling H, Eggert C, Harréus U and Gires O: EpCAM regulates cell cycle progression via control of cyclin D1 expression. Oncogene. 32:641–650. 2013. View Article : Google Scholar
19	Münz M, Kieu C, Mack B, Schmitt B, Zeidler R and Gires O: The carcinoma-associated antigen EpCAM upregulates c-myc and induces cell proliferation. Oncogene. 23:5748–5758. 2004. View Article : Google Scholar : PubMed/NCBI
20	Litvinov SV, Balzar M, Winter MJ, Bakker HA, Bruijn IH, Prins F, Fleuren GJ and Warnaar SO: Epithelial cell adhesion molecule (Ep-CAM) modulates cell-cell interactions mediated by classic cadherins. J Cell Biol. 139:1337–1348. 1997. View Article : Google Scholar
21	Lin CW, Liao MY, Lin WW, Wang YP, Lu TY and Wu HC: Epithelial cell adhesion molecule regulates tumor initiation and tumorigenesis via activating reprogramming factors and epithelial-mesenchymal transition gene expression in colon cancer. J Biol Chem. 287:39449–39459. 2012. View Article : Google Scholar : PubMed/NCBI
22	Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009. View Article : Google Scholar : PubMed/NCBI
23	Philip R, Heiler S, Mu W, Buchler MW, Zoller M and Thuma F: Claudin-7 promotes the epithelial-mesenchymal transition in human colorectal cancer. Oncotarget. 6:2046–2063. 2015. View Article : Google Scholar
24	Medici D, Hay ED and Olsen BR: Snail and Slug promote epithelial-mesenchymal transition through beta-catenin-T-cell factor-4-dependent expression of transforming growth factor-beta3. Mol Biol Cell. 19:4875–4887. 2008. View Article : Google Scholar : PubMed/NCBI
25	Garg M: Epithelial-mesenchymal transition - activating transcription factors - multifunctional regulators in cancer. World J Stem Cells. 5:188–195. 2013. View Article : Google Scholar : PubMed/NCBI
26	Bokemeyer C: Catumaxomab - trifunctional anti-EpCAM antibody used to treat malignant ascites. Expert Opin Biol Ther. 10:1259–1269. 2010. View Article : Google Scholar : PubMed/NCBI
27	MacDonald GC, Rasamoelisolo M, Entwistle J, Cuthbert W, Kowalski M, Spearman MA and Glover N: A phase I clinical study of intratumorally administered VB4-845, an anti-epithelial cell adhesion molecule recombinant fusion protein, in patients with squamous cell carcinoma of the head and neck. Med Oncol. 26:257–264. 2009. View Article : Google Scholar
28	Kowalski M, Guindon J, Brazas L, Moore C, Entwistle J, Cizeau J, Jewett MAS and MacDonald GC: A phase II study of oportuzumab monatox: An immunotoxin therapy for patients with noninvasive urothelial carcinoma in situ previously treated with bacillus Calmette-Guerin. J Urol. 188:1712–1718. 2012. View Article : Google Scholar : PubMed/NCBI
29	Ogawa K, Tanaka S, Matsumura S, Murakata A, Ban D, Ochiai T, Irie T, Kudo A, Nakamura N, Tanabe M, et al: EpCAM-targeted therapy for human hepatocellular carcinoma. Ann Surg Oncol. 21:1314–1322. 2014. View Article : Google Scholar