Eps8 regulates cellular proliferation and migration of breast cancer

Chen,Cheng; Liang,Zhongheng; Huang,Wenhuan; Li,Xinxin; Zhou,Fangliang; Hu,Xiang; Han,Mei; Ding,Xiaofeng; Xiang,Shuanglin

doi:10.3892/ijo.2014.2710

Eps8 regulates cellular proliferation and migration of breast cancer

Authors:
- Cheng Chen
- Zhongheng Liang
- Wenhuan Huang
- Xinxin Li
- Fangliang Zhou
- Xiang Hu
- Mei Han
- Xiaofeng Ding
- Shuanglin Xiang
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Affiliations: Key Laboratory of Protein Chemistry and Development Biology of State Education Ministry of China, College of Life Science, Hunan Normal University, Changsha, P.R. China, College of Basic Medical Sciences, Hunan University of Chinese Medicine, Changsha, P.R. China
Published online on: October 17, 2014 https://doi.org/10.3892/ijo.2014.2710
Pages: 205-214

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Abstract

The role of Eps8 in human breast cancer was studied, and we found that Eps8 was overexpressed in >60% of human breast cancer samples compared with adjacent normal breast tissues by immunohistochemical analysis. Eps8 was highly expressed in the highly invasive breast cancer cell line MDA-MB‑231 compared with the weakly invasive breast cancer cell lines MCF7 and MDA-MB‑468. MCF7 cell line stably expressing Eps8 was established by G418 screening, and the ectopic expression of Eps8 enhanced MCF7 breast cancer cell growth and survival as assessed by MTT analysis, cell viability and liquid colony formation, whereas the lentiviral expression of Eps8 shRNA in MDA-MB‑231 cells resulted in a significant reduction in cellular growth and proliferation in vitro and in vivo. Furthermore, Eps8 knockdown inhibited breast cancer cell migration in wound healing assays, decreased the number and size of EGF-induced filopodia and increased the sensitivity of breast cancer cells to cisplatin analyzed by MTT assays. Eps8 knockdown decreased the levels of phosphorylated extracellular signal-regulated protein kinase (ERK) and MMP9 but increased p53. Moreover, Eps8 knockdown suppressed a partial EMT-like transition and showed a significant increase in E-cadherin and decrease in N-cadherin and vimentin. These results suggest that Eps8 is overexpressed in human breast cancers, possibly by regulating ERK signaling, MMP9, p53 and EMT-like transition to affect breast cancer cell growth, migration and invasion. Therefore, Eps8 might represent a novel potential target in human breast cancer therapy.

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1	Bray F, McCarron P and Parkin DM: The changing global patterns of female breast cancer incidence and mortality. Breast Cancer Res. 6:229–239. 2004. View Article : Google Scholar : PubMed/NCBI
2	Gupta GP and Massague J: Cancer metastasis: building a framework. Cell. 127:679–695. 2006. View Article : Google Scholar : PubMed/NCBI
3	Dahiya R and Deng G: Molecular prognostic markers in breast cancer. Breast Cancer Res Treat. 52:185–200. 1998. View Article : Google Scholar : PubMed/NCBI
4	Ross JS and Fletcher JA: The HER-2/neu oncogene in breast cancer: prognostic factor, predictive factor, and target for therapy. Oncologist. 3:237–252. 1998.
5	Sgouros J, Galani E, Gonos E, et al: Correlation of nm23-H1 gene expression with clinical outcome in patients with advanced breast cancer. In Vivo. 21:519–522. 2007.PubMed/NCBI
6	Barnes DM and Gillett CE: Cyclin D1 in breast cancer. Breast Cancer Res Treat. 52:1–15. 1998. View Article : Google Scholar
7	Wang H, Teh MT, Ji Y, et al: EPS8 upregulates FOXM1 expression, enhancing cell growth and motility. Carcinogenesis. 31:1132–1141. 2010. View Article : Google Scholar : PubMed/NCBI
8	Chen YJ, Shen MR, Chen YJ, Maa MC and Leu TH: Eps8 decreases chemosensitivity and affects survival of cervical cancer patients. Mol Cancer Ther. 7:1376–1385. 2008. View Article : Google Scholar : PubMed/NCBI
9	Maa MC, Lee JC, Chen YJ, et al: Eps8 facilitates cellular growth and motility of colon cancer cells by increasing the expression and activity of focal adhesion kinase. J Biol Chem. 282:19399–19409. 2007. View Article : Google Scholar : PubMed/NCBI
10	Wang H, Patel V, Miyazaki H, Gutkind JS and Yeudall WA: Role for EPS8 in squamous carcinogenesis. Carcinogenesis. 30:165–174. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zhang W, Wang L, Liu Y, et al: Structure of human lanthionine synthetase C-like protein 1 and its interaction with Eps8 and glutathione. Genes Dev. 23:1387–1392. 2009. View Article : Google Scholar : PubMed/NCBI
12	Welsch T, Endlich K, Giese T, Buchler MW and Schmidt J: Eps8 is increased in pancreatic cancer and required for dynamic actin-based cell protrusions and intercellular cytoskeletal organization. Cancer Lett. 255:205–218. 2007. View Article : Google Scholar
13	Ding X, Zhou F, Wang F, et al: Eps8 promotes cellular growth of human malignant gliomas. Oncol Rep. 29:697–703. 2013.PubMed/NCBI
14	Kang H, Wilson CS, Harvey RC, et al: Gene expression profiles predictive of outcome and age in infant acute lymphoblastic leukemia: a Children’s Oncology Group study. Blood. 119:1872–1881. 2012. View Article : Google Scholar : PubMed/NCBI
15	Matoskova B, Wong WT, Salcini AE, Pelicci PG and Di Fiore PP: Constitutive phosphorylation of eps8 in tumor cell lines: relevance to malignant transformation. Mol Cell Biol. 15:3805–3812. 1995.PubMed/NCBI
16	Chu PY, Liou JH, Lin YM, et al: Expression of Eps8 correlates with poor survival in oral squamous cell carcinoma. Asia Pac J Clin Oncol. 8:e77–e81. 2012. View Article : Google Scholar : PubMed/NCBI
17	Yang TP, Chiou HL, Maa MC and Wang CJ: Mithramycin inhibits human epithelial carcinoma cell proliferation and migration involving downregulation of Eps8 expression. Chem Biol Interact. 183:181–186. 2010. View Article : Google Scholar : PubMed/NCBI
18	Gorsic LK, Stark AL, Wheeler HE, et al: EPS8 inhibition increases cisplatin sensitivity in lung cancer cells. PLoS One. 8:e822202013. View Article : Google Scholar : PubMed/NCBI
19	Abdel-Rahman WM, Ruosaari S, Knuutila S and Peltomaki P: Differential roles of EPS8 in carcinogenesis: loss of protein expression in a subset of colorectal carcinoma and adenoma. World J Gastroenterol. 18:3896–3903. 2012. View Article : Google Scholar : PubMed/NCBI
20	Li YH, Xue TY, He YZ and Du JW: Novel oncoprotein EPS8: a new target for anticancer therapy. Future Oncol. 9:1587–1594. 2013. View Article : Google Scholar : PubMed/NCBI
21	Yao J, Weremowicz S, Feng B, et al: Combined cDNA array comparative genomic hybridization and serial analysis of gene expression analysis of breast tumor progression. Cancer Res. 66:4065–4078. 2006. View Article : Google Scholar : PubMed/NCBI
22	Mangravite LM, Lipschutz JH, Mostov KE and Giacomini KM: Localization of GFP-tagged concentrative nucleoside transporters in a renal polarized epithelial cell line. Am J Physiol Renal Physiol. 280:F879–F885. 2001.PubMed/NCBI
23	Schmid JA, Birbach A, Hofer-Warbinek R, et al: Dynamics of NF kappa B and Ikappa Balpha studied with green fluorescent protein (GFP) fusion proteins. Investigation of GFP-p65 binding to DNa by fluorescence resonance energy transfer. J Biol Chem. 275:17035–17042. 2000. View Article : Google Scholar
24	Lois C, Hong EJ, Pease S, Brown EJ and Baltimore D: Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science. 295:868–872. 2002. View Article : Google Scholar : PubMed/NCBI
25	Ding X, Yang Z, Zhou F, et al: Transcription factor AP-2alpha regulates acute myeloid leukemia cell proliferation by influencing Hoxa gene expression. Int J Biochem Cell Biol. 45:1647–1656. 2013. View Article : Google Scholar : PubMed/NCBI
26	Pan CC, Bloodworth JC, Mythreye K and Lee NY: Endoglin inhibits ERK-induced c-Myc and cyclin D1 expression to impede endothelial cell proliferation. Biochem Biophys Res Commun. 424:620–623. 2012. View Article : Google Scholar : PubMed/NCBI
27	Ridley AJ: Rho GTPases and cell migration. J Cell Sci. 114:2713–2722. 2001.PubMed/NCBI
28	Scita G, Tenca P, Areces LB, et al: An effector region in Eps8 is responsible for the activation of the Rac-specific GEF activity of Sos-1 and for the proper localization of the Rac-based actin-polymerizing machine. J Cell Biol. 154:1031–1044. 2001. View Article : Google Scholar : PubMed/NCBI
29	Scita G, Nordstrom J, Carbone R, et al: EPS8 and E3B1 transduce signals from Ras to Rac. Nature. 401:290–293. 1999. View Article : Google Scholar : PubMed/NCBI
30	Chen H, Wu X, Pan ZK and Huang S: Integrity of SOS1/EPS8/ ABI1 tri-complex determines ovarian cancer metastasis. Cancer Res. 70:9979–9990. 2010. View Article : Google Scholar : PubMed/NCBI
31	Hecquet C, Lefevre G, Valtink M, Engelmann K and Mascarelli F: Activation and role of MAP kinase-dependent pathways in retinal pigment epithelial cells: ERK and RPE cell proliferation. Invest Ophthalmol Vis Sci. 43:3091–3098. 2002.PubMed/NCBI
32	Chang F, Steelman LS, Lee JT, et al: Signal transduction mediated by the Ras/Raf/MEK/ERK pathway from cytokine receptors to transcription factors: potential targeting for therapeutic intervention. Leukemia. 17:1263–1293. 2003. View Article : Google Scholar
33	McCawley LJ, Li S, Wattenberg EV and Hudson LG: Sustained activation of the mitogen-activated protein kinase pathway. A mechanism underlying receptor tyrosine kinase specificity for matrix metalloproteinase-9 induction and cell migration. J Biol Chem. 274:4347–4353. 1999. View Article : Google Scholar
34	Oren M: Decision making by p53: life, death and cancer. Cell Death Differ. 10:431–442. 2003. View Article : Google Scholar : PubMed/NCBI
35	Thiery JP: Epithelial-mesenchymal transitions in development and pathologies. Curr Opin Cell Biol. 15:740–746. 2003. View Article : Google Scholar : PubMed/NCBI
36	Grunert S, Jechlinger M and Beug H: Diverse cellular and molecular mechanisms contribute to epithelial plasticity and metastasis. Nat Rev Mol Cell Biol. 4:657–665. 2003. View Article : Google Scholar : PubMed/NCBI
37	Onder TT, Gupta PB, Mani SA, et al: Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 68:3645–3654. 2008. View Article : Google Scholar : PubMed/NCBI
38	Tanaka H, Kono E, Tran CP, et al: Monoclonal antibody targeting of N-cadherin inhibits prostate cancer growth, metastasis and castration resistance. Nat Med. 16:1414–1420. 2010. View Article : Google Scholar : PubMed/NCBI
39	Lang SH, Hyde C, Reid IN, et al: Enhanced expression of vimentin in motile prostate cell lines and in poorly differentiated and metastatic prostate carcinoma. Prostate. 52:253–263. 2002. View Article : Google Scholar : PubMed/NCBI