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Open Access 01-12-2012 | Research article

Nuclear translocation of the cytoplasmic domain of HB-EGF induces gastric cancer invasion

Authors: Takaya Shimura, Michihiro Yoshida, Shinji Fukuda, Masahide Ebi, Yoshikazu Hirata, Tsutomu Mizoshita, Satoshi Tanida, Hiromi Kataoka, Takeshi Kamiya, Shigeki Higashiyama, Takashi Joh

Published in: BMC Cancer | Issue 1/2012

Abstract

Background

Membrane-anchored heparin-binding epidermal growth factor-like growth factor (proHB-EGF) yields soluble HB-EGF, which is an epidermal growth factor receptor (EGFR) ligand, and a carboxy-terminal fragment of HB-EGF (HB-EGF-CTF) after ectodomain shedding. We previously reported that HB-EGF-CTF and unshed proHB-EGF which has the cytoplasmic domain of proHB-EGF (HB-EGF-C), translocate from the plasma membrane to the nucleus and regulate cell cycle after shedding stimuli. However, the significance of nuclear exported HB-EGF-C in human gastric cancer is unclear.

Methods

We investigated the relationship between intracellular localization of HB-EGF-C and clinical outcome in 96 gastric cancer patients treated with gastrectomy. Moreover, we established stable gastric cancer cell lines overexpressing wild-type HB-EGF (wt-HB-EGF) and mutated HB-EGF (HB-EGF-mC), which prevented HB-EGF-C nuclear translocation after shedding. Cell motility between these 2 gastric cancer cell lines was investigated using a transwell invasion assay and a wound healing assay.

Results

Of the 96 gastric cancer cases, HB-EGF-C immunoreactivity was detected in both the nucleus and cytoplasm in 19 cases (19.8 %) and in the cytoplasm only in 25 cases (26.0 %). The nuclear immunoreactivity of HB-EGF-C was significantly increased in stage pT3/4 tumors compared with pT1/2 tumors (T1/2 vs. T3/4: 11.1 % vs. 36.4 %, P < 0.01). The growth of wt-HB-EGF- and HB-EGF-mC-expressing cells significantly increased compared with control cells, but the growth of HB-EGF-mC-expressing cells was significantly decreased compared with wt-HB-EGF-expressing cells. Gastric cancer cell invasion obviously increased in wt-HB-EGF-expressing cells, but invasion in HB-EGF-mC-expressing cells showed a slight increase compared with control cells. Moreover, wt-HB-EGF overexpression increased the effectiveness of wound healing, but had no significant effect in HB-EGF-mC-expressing cells.

Conclusions

Both the function of HB-EGF as an EGFR ligand and a novel signal for HB-EGF-C nuclear translocation induce gastric cancer growth, whereas HB-EGF-C nuclear translocation independently plays a critical role in gastric cancer invasion. The present study demonstrated that HB-EGF-C nuclear translocation might be crucial in gastric cancer invasion. HB-EGF-C nuclear translocation may offer a prognostic marker and a new molecular target for gastric cancer therapy.

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International Agency for Research on Cancer GLOBOCAN 2008: http://globocan.iarc.fr/factsheets/populations/factsheet.asp?uno=900 (last accessed on December 16, 2011)

Bang YJ, Van Cutsem E, Feyereislova A, Chung HC, Shen L, Sawaki A, Lordick F, Ohtsu A, Omuro Y, Satoh T, et al: Trastuzumab in combination with chemotherapy versus chemotherapy alone for treatment of HER2-positive advanced gastric or gastro-oesophageal junction cancer (ToGA): a phase 3, open-label, randomised controlled trial. Lancet. 2010, 376: 687-697. 10.1016/S0140-6736(10)61121-X.CrossRefPubMed

Higashiyama S, Iwabuki H, Morimoto C, Hieda M, Inoue H, Matsushita N: Membrane-anchored growth factors, the epidermal growth factor family: beyond receptor ligands. Cancer Sci. 2008, 99: 214-220. 10.1111/j.1349-7006.2007.00676.x.CrossRefPubMed

Normanno N, Bianco C, De Luca A, Maiello MR, Salomon DS: Target-based agents against ErbB receptors and their ligands: a novel approach to cancer treatment. Endocr Relat Cancer. 2003, 10: 1-21. 10.1677/erc.0.0100001.CrossRefPubMed

Salomon DS, Brandt R, Ciardiello F, Normanno N: Epidermal growth factor-related peptides and their receptors in human malignancies. Crit Rev Oncol Hematol. 1995, 19: 183-232. 10.1016/1040-8428(94)00144-I.CrossRefPubMed

Kinugasa Y, Hieda M, Hori M, Higashiyama S: The carboxyl-terminal fragment of pro-HB-EGF reverses Bcl6-mediated gene repression. J Biol Chem. 2007, 282: 14797-14806. 10.1074/jbc.M611036200.CrossRefPubMed

Nanba D, Mammoto A, Hashimoto K, Higashiyama S: Proteolytic release of the carboxy-terminal fragment of proHB-EGF causes nuclear export of PLZF. J Cell Biol. 2003, 163: 489-502. 10.1083/jcb.200303017.CrossRefPubMedPubMedCentral

Hirata Y, Ogasawara N, Sasaki M, Mizushima T, Shimura T, Mizoshita T, Mori Y, Kubota E, Wada T, Tanida S, et al: BCL6 degradation caused by the interaction with the C-terminus of pro-HB-EGF induces cyclin D2 expression in gastric cancers. Br J Cancer. 2009, 100: 1320-1329. 10.1038/sj.bjc.6605010.CrossRefPubMedPubMedCentral

Shimura T, Kataoka H, Ogasawara N, Kubota E, Sasaki M, Tanida S, Joh T: Suppression of proHB-EGF carboxy-terminal fragment nuclear translocation: a new molecular target therapy for gastric cancer. Clin Cancer Res. 2008, 14: 3956-3965. 10.1158/1078-0432.CCR-07-4794.CrossRefPubMed

10.

Hieda M, Isokane M, Koizumi M, Higashi C, Tachibana T, Shudou M, Taguchi T, Hieda Y, Higashiyama S: Membrane-anchored growth factor, HB-EGF, on the cell surface targeted to the inner nuclear membrane. J Cell Biol. 2008, 180: 763-769. 10.1083/jcb.200710022.CrossRefPubMedPubMedCentral

11.

Tokumaru S, Higashiyama S, Endo T, Nakagawa T, Miyagawa JI, Yamamori K, Hanakawa Y, Ohmoto H, Yoshino K, Shirakata Y, et al: Ectodomain shedding of epidermal growth factor receptor ligands is required for keratinocyte migration in cutaneous wound healing. J Cell Biol. 2000, 151: 209-220. 10.1083/jcb.151.2.209.CrossRefPubMedPubMedCentral

12.

Marikovsky M, Breuing K, Liu PY, Eriksson E, Higashiyama S, Farber P, Abraham J, Klagsbrun M: Appearance of heparin-binding EGF-like growth factor in wound fluid as a response to injury. Proc Natl Acad Sci USA. 1993, 90: 3889-3893. 10.1073/pnas.90.9.3889.CrossRefPubMedPubMedCentral

13.

Hynes NE, Lane HA: ERBB receptors and cancer: the complexity of targeted inhibitors. Nat Rev Cancer. 2005, 5: 341-354. 10.1038/nrc1609.CrossRefPubMed

14.

Nanba D, Inoue H, Shigemi Y, Shirakata Y, Hashimoto K, Higashiyama S: An intermediary role of proHB-EGF shedding in growth factor-induced c-Myc gene expression. J Cell Physiol. 2008, 214: 465-473. 10.1002/jcp.21233.CrossRefPubMed

15.

Iwamoto R, Yamazaki S, Asakura M, Takashima S, Hasuwa H, Miyado K, Adachi S, Kitakaze M, Hashimoto K, Raab G, et al: Heparin-binding EGF-like growth factor and ErbB signaling is essential for heart function. Proc Natl Acad Sci USA. 2003, 100: 3221-3226. 10.1073/pnas.0537588100.CrossRefPubMedPubMedCentral

16.

Wang F, Liu R, Lee SW, Sloss CM, Couget J, Cusack JC: Heparin-binding EGF-like growth factor is an early response gene to chemotherapy and contributes to chemotherapy resistance. Oncogene. 2007, 26: 2006-2016. 10.1038/sj.onc.1209999.CrossRefPubMed

17.

Ongusaha PP, Kwak JC, Zwible AJ, Macip S, Higashiyama S, Taniguchi N, Fang L, Lee SW: HB-EGF is a potent inducer of tumor growth and angiogenesis. Cancer Res. 2004, 64: 5283-5290. 10.1158/0008-5472.CAN-04-0925.CrossRefPubMed

18.

Inui Y, Higashiyama S, Kawata S, Tamura S, Miyagawa J, Taniguchi N, Matsuzawa Y: Expression of heparin-binding epidermal growth factor in human hepatocellular carcinoma. Gastroenterology. 1994, 107: 1799-1804.CrossRefPubMed

19.

Kobrin MS, Funatomi H, Friess H, Buchler MW, Stathis P, Korc M: Induction and expression of heparin-binding EGF-like growth factor in human pancreatic cancer. Biochem Biophys Res Commun. 1994, 202: 1705-1709. 10.1006/bbrc.1994.2131.CrossRefPubMed

20.

Murayama Y, Miyagawa J, Shinomura Y, Kanayama S, Isozaki K, Yamamori K, Mizuno H, Ishiguro S, Kiyohara T, Miyazaki Y, et al: Significance of the association between heparin-binding epidermal growth factor-like growth factor and CD9 in human gastric cancer. Int J Cancer. 2002, 98: 505-513. 10.1002/ijc.10198.CrossRefPubMed

21.

Naef M, Yokoyama M, Friess H, Buchler MW, Korc M: Co-expression of heparin-binding EGF-like growth factor and related peptides in human gastric carcinoma. Int J Cancer. 1996, 66: 315-321. 10.1002/(SICI)1097-0215(19960503)66:3<315::AID-IJC8>3.0.CO;2-1.CrossRefPubMed

22.

Kopp R, Rothbauer E, Ruge M, Arnholdt H, Spranger J, Muders M, Pfeiffer DG, Schildberg FW, Pfeiffer A: Clinical implications of the EGF receptor/ligand system for tumor progression and survival in gastrointestinal carcinomas: evidence for new therapeutic options. Recent Results Cancer Res. 2003, 162: 115-132. 10.1007/978-3-642-59349-9_10.CrossRefPubMed

23.

Yagi H, Miyamoto S, Tanaka Y, Sonoda K, Kobayashi H, Kishikawa T, Iwamoto R, Mekada E, Nakano H: Clinical significance of heparin-binding epidermal growth factor-like growth factor in peritoneal fluid of ovarian cancer. Br J Cancer. 2005, 92: 1737-1745. 10.1038/sj.bjc.6602536.CrossRefPubMedPubMedCentral

24.

Yasumoto K, Yamada T, Kawashima A, Wang W, Li Q, Shterev DI, Tacheuchi S, Mouri H, Yamashita K, Ohtsubo K, Yano S: The EGFR ligands Amphiregulin and Heparin-binding EGF-like growth factor promote peritoneal carcinomatosis in CXCR4-expressing gastric cancer. Clin Cancer Res. 2011, 17: 3619-3630. 10.1158/1078-0432.CCR-10-2475.CrossRefPubMed

25.

Wang F, Sloss C, Zhang X, Lee SW, Cusack JC: Membrane-bound heparin-binding epidermal growth factor like growth factor regulates E-cadherin expression in pancreatic carcinoma cells. Cancer Res. 2007, 67: 8486-8493. 10.1158/0008-5472.CAN-07-0498.CrossRefPubMed

26.

Singh AB, Tsukada T, Zent R, Harris RC: Membrane-associated HB-EGF modulates HGF-induced cellular responses in MDCK cells. J Cell Sci. 2004, 117: 1365-1379. 10.1242/jcs.01037.CrossRefPubMed

27.

Kramer C, Klasmeyer K, Bojar H, Schulz WA, Ackermann R, Grimm MO: Heparin-binding epidermal growth factor-like growth factor isoforms and epidermal growth factor receptor/ErbB1 expression in bladder cancer and their relation to clinical outcome. Cancer. 2007, 109: 2016-2024. 10.1002/cncr.22627.CrossRefPubMed

28.

Adam RM, Danciu T, McLellan DL, Borer JG, Lin J, Zurakowski D, Weinstein MH, Rajjayabun PH, Mellon JK, Freeman MR: A nuclear form of the heparin-binding epidermal growth factor-like growth factor precursor is a feature of aggressive transitional cell carcinoma. Cancer Res. 2003, 63: 484-490.PubMed

29.

Sobin L, Gospodarowicz M, Wittekind C: TNM classification of malignant tumours. Edited by: Hoboken NJ. 2009, John Wiley & Sons, Inc, 7

30.

Goishi K, Higashiyama S, Klagsbrun M, Nakano N, Umata T, Ishikawa M, Mekada E, Taniguchi N: Phorbol ester induces the rapid processing of cell surface heparin-binding EGF-like growth factor: conversion from juxtacrine to paracrine growth factor activity. Mol Biol Cell. 1995, 6: 967-980.CrossRefPubMedPubMedCentral

The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/12/205/prepub

Title: Nuclear translocation of the cytoplasmic domain of HB-EGF induces gastric cancer invasion
Authors: Takaya Shimura
Michihiro Yoshida
Shinji Fukuda
Masahide Ebi
Yoshikazu Hirata
Tsutomu Mizoshita
Satoshi Tanida
Hiromi Kataoka
Takeshi Kamiya
Shigeki Higashiyama
Takashi Joh
Publication date: 01-12-2012
Publisher: BioMed Central
Published in: BMC Cancer / Issue 1/2012
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/1471-2407-12-205

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