14-3-3ε is a nuclear matrix protein, and its altered expression and localization are associated with curcumin-induced apoptosis of MG‑63 cells

Lu,Kun; Rui,Gang; Liu,Fan; Yang,Ling; Deng,Xiaoling; Shi,Songlin; Li,Qifu

doi:10.3892/ol.2017.7283

14-3-3ε is a nuclear matrix protein, and its altered expression and localization are associated with curcumin-induced apoptosis of MG‑63 cells

Authors:
- Kun Lu
- Gang Rui
- Fan Liu
- Ling Yang
- Xiaoling Deng
- Songlin Shi
- Qifu Li
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Affiliations: Department of Basic Medicine, Medical College of Xiamen University, Xiamen, Fujian 361102, P.R. China, Department of Orthopedics, The First Affiliated Hospital of Xiamen University, Xiamen, Fujian 361003, P.R. China
Published online on: October 30, 2017 https://doi.org/10.3892/ol.2017.7283
Pages: 338-346
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The 14-3-3 protein family may regulates protein interaction, transportation and cellular localization. The regulatory role of 14‑3‑3ε is influenced by its altered localization. In the present study, human osteosarcoma MG‑63 cells were treated with curcumin to induce apoptosis. Subsequently, the altered expression and localization of 14‑3‑3ε and its co‑localization with other apoptosis‑associated proteins during apoptosis was investigated. Analysis of nuclear matrix proteins (NMPs), using two‑dimensional gel electrophoresis with matrix‑assisted laser‑desorption/ionization time‑of‑flight mass spectrometry, revealed that 14‑3‑3ε existed on the nuclear matrix of MG‑63 cells, and its expression was decreased compared with that in control cells following curcumin treatment. In addition, western blot analysis validated that the expression level of 14‑3‑3ε was downregulated during curcumin‑induced apoptosis of MG‑63 cells compared with that in control cells. Using immunofluorescence labeling, it was observed that 14‑3‑3ε was located on the nuclear matrix of MG‑63 cells and the distribution of 14‑3‑3ε on the nuclear matrix was decreased following treatment with curcumin, compared with that in control cells. Double immunofluorescence staining and laser‑scanning confocal microscopy demonstrated that 14‑3‑3ε was co‑localized with B‑cell lymphoma‑2 (Bcl‑2), Bcl‑2‑associated‑X protein, p53 and c‑FOS transcription factor in MG‑63 cells. Furthermore, following treatment with curcumin, these co‑localization regions were decreased. The results of the present study revealed that 14‑3‑3ε is an NMP in MG‑63 cells, and its altered expression and co‑localization with apoptosis‑associated proteins indicated an important function of 14‑3‑3ε in apoptosis of MG‑63 cells. Additional studies are required to investigate the results of the present study.

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1	Morrison DK: The 14-3-3 proteins: Integrators of diverse signaling cues that impact cell fate and cancer development. Trends Cell Biol. 19:16–23. 2009. View Article : Google Scholar : PubMed/NCBI
2	van Hemert MJ, Steensma HY and van Heusden GP: 14-3-3 proteins: Key regulators of cell division, signalling and apoptosis. Bioessays. 23:936–946. 2001. View Article : Google Scholar : PubMed/NCBI
3	Kosaka Y, Cieslik KA, Li L, Lezin G, Maguire CT, Saijoh Y, Toyo-oka K, Gambello MJ, Vatta M, Wynshaw-Boris A, et al: 14-3-3ε plays a role in cardiac ventricular compaction by regulating the cardiomyocyte cell cycle. Mol Cell Biol. 32:5089–5102. 2012. View Article : Google Scholar : PubMed/NCBI
4	Cui C, Ren X, Liu D, Deng X, Qin X, Zhao X, Wang E and Yu B: 14-3-3 epsilon prevents G2/M transition of fertilized mouse eggs by binding with CDC25B. BMC Dev Biol. 14:332014. View Article : Google Scholar : PubMed/NCBI
5	Nagappan A, Park HS, Park KI, Hong GE, Yumnam S, Lee HJ, Kim MK, Kim EH, Lee WS, Lee WJ, et al: Helicobacter pylori infection combined with DENA revealed altered expression of p53 and 14-3-3 isoforms in Gulo-/- mice. Chem Biol Interact. 206:143–152. 2013. View Article : Google Scholar : PubMed/NCBI
6	Zhong J, Kong X, Zhang H, Yu C, Xu Y, Kang J, Yu H, Yi H, Yang X and Sun L: Inhibition of CLIC4 enhances autophagy and triggers mitochondrial and ER stress-induced apoptosis in human glioma U251 cells under starvation. PLoS One. 7:e393782012. View Article : Google Scholar : PubMed/NCBI
7	Liu Y, Song F, Wu WK, He M, Zhao L, Sun X, Li H, Jiang Y, Yang Y and Peng K: Triptolide inhibits colon cancer cell proliferation and induces cleavage and translocation of 14-3-3 epsilon. Cell Biochem Funct. 30:271–278. 2012. View Article : Google Scholar : PubMed/NCBI
8	Fong WH, Tsai HD, Chen YC, Wu JS and Lin TN: Anti-apoptotic actions of PPAR-gamma against ischemic stroke. Mol Neurobiol. 41:180–186. 2010. View Article : Google Scholar : PubMed/NCBI
9	Wang X, Grammatikakis N, Siganou A and Calderwood SK: Regulation of molecular chaperone gene transcription involves the serine phosphorylation, 14-3-3 epsilon binding, and cytoplasmic sequestration of heat shock factor 1. Mol Cell Biol. 23:6013–6026. 2003. View Article : Google Scholar : PubMed/NCBI
10	Leal MF, Calcagno DQ, Demachki S, Assumpcão PP, Chammas R, Burbano RR and Mde A Smith: Clinical implication of 14-3-3 epsilon expression in gastric cancer. World J Gastroenterol. 18:1531–1537. 2012. View Article : Google Scholar : PubMed/NCBI
11	Ruiz E, sparza-Garrido R, Velázquez-Flores MÁ, Diegopérez-Ramirez J, López-Aguilar E, Siordia-Reyes G, Hernández-Ortiz M, Martinez-Batallar AG, Encarnación-Guevara S, Salamanca-Gómez F and Arenas-Aranda DJ: A proteomic approach of pediatric astrocytomas: MiRNAs and network insight. J Proteomics. 94:162–175. 2013. View Article : Google Scholar : PubMed/NCBI
12	Ko BS, Jan YJ, Chang TC, Liang SM, Chen SC, Liu TA, Wu YM, Wang J and Liou JY: Upregulation of focal adhesion kinase by 14-3-3ε via NFκB activation in hepatocellular carcinoma. Anticancer Agents Med Chem. 13:555–562. 2013. View Article : Google Scholar : PubMed/NCBI
13	Ko BS, Chang TC, Hsu C, Chen YC, Shen TL, Chen SC, Wang J, Wu KK, Jan YJ and Liou JY: Overexpression of 14-3-3ε predicts tumour metastasis and poor survival in hepatocellular carcinoma. Histopathology. 58:705–711. 2011. View Article : Google Scholar : PubMed/NCBI
14	Liang S, Xu Y, Shen G, Liu Q, Zhao X, Xu Z, Xie X, Gong F, Li R and Wei Y: Quantitative protein expression profiling of 14-3-3 isoforms in human renal carcinoma shows 14-3-3 epsilon is involved in limitedly increasing renal cell proliferation. Electrophoresis. 30:4152–4162. 2009. View Article : Google Scholar : PubMed/NCBI
15	Cimino D, Fuso L, Sfiligoi C, Biglia N, Ponzone R, Maggiorotto F, Russo G, Cicatiello L, Weisz A, Taverna D, et al: Identification of new genes associated with breast cancer progression by gene expression analysis of predefined sets of neoplastic tissues. Int J Cancer. 123:1327–1338. 2008. View Article : Google Scholar : PubMed/NCBI
16	Porter GW, Khuri FR and Fu H: Dynamic 14-3-3/client protein interactions integrate survival and apoptotic pathways. Semin Cancer Biol. 16:193–202. 2006. View Article : Google Scholar : PubMed/NCBI
17	Zha J, Harada H, Yang E, Jockel J and Korsmeyer SJ: Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell. 87:619–628. 1996. View Article : Google Scholar : PubMed/NCBI
18	Nomura M, Shimizu S, Sugiyama T, Narita M, Ito T, Matsuda H and Tsujimoto Y: 14-3-3 Interacts directly with and negatively regulates pro-apoptotic Bax. J Biol Chem. 278:2058–2065. 2003. View Article : Google Scholar : PubMed/NCBI
19	Zhang L, Chen J and Fu H: Suppression of apoptosis signal-regulating kinase 1-induced cell death by 14-3-3 proteins. Proc Natl Acad Sci USA. 96:pp. 8511–8515. 1999; View Article : Google Scholar : PubMed/NCBI
20	Brunet A, Kanai F, Stehn J, Xu J, Sarbassova D, Frangioni JV, Dalal SN, DeCaprio JA, Greenberg ME and Yaffe MB: 14-3-3 transits to the nucleus and participates in dynamic nucleocytoplasmic transport. J Cell Biol. 156:817–828. 2002. View Article : Google Scholar : PubMed/NCBI
21	Fey EG, Wan KM and Penman S: Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: Three-dimensional organization and protein composition. J Cell Biol. 98:1973–1984. 1984. View Article : Google Scholar : PubMed/NCBI
22	Gerner C, Seelos C and Sauermann G: Alteration of nuclear matrix protein composition during apoptosis in rat embryo cells. Exp Cell Res. 238:472–480. 1998. View Article : Google Scholar : PubMed/NCBI
23	Michishita E, Kurahashi T, Suzuki T, Fukuda M, Fujii M, Hirano H and Ayusawa D: Changes in nuclear matrix proteins during the senescence-like phenomenon induced by 5-chlorodeoxyuridine in HeLa cells. Exp Gerontol. 37:885–890. 2002. View Article : Google Scholar : PubMed/NCBI
24	Jing GJ, Xu DH, Shi SL, Li QF, Wang SY, Wu FY and Kong HY: Aberrant expression of nuclear matrix proteins during HMBA-induced differentiation of gastric cancer cells. World J Gastroenterol. 16:2176–2182. 2010. View Article : Google Scholar : PubMed/NCBI
25	Shevchenko A, Wilm M, Vorm O and Mann M: Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem. 68:850–858. 1996. View Article : Google Scholar : PubMed/NCBI
26	Boutet E, Lieberherr D, Tognolli M, Schneider M, Bansal P, Bridge AJ, Poux S, Bougueleret L and Xenarios I: UniProtKB/Swiss-Prot, the manually annotated section of the UniProt KnowledgeBase: How to use the entry view. Methods Mol Biol. 1374:23–54. 2016. View Article : Google Scholar : PubMed/NCBI
27	NCBI Resource Coordinators, . Database resources of the national center for biotechnology information. Nucleic Acids Res. 45:D12–D17. 2017. View Article : Google Scholar : PubMed/NCBI
28	Li QF: Effect of retinoic acid on the changes of nuclear matrix in termediate filament system in gastric carcinoma cells. World J Gastroenterol. 5:417–420. 1999. View Article : Google Scholar : PubMed/NCBI
29	Poüs C, Klipfel L and Baillet A: Cancer-related functions and subcellular localizations of septins. Front Cell Dev Biol. 4:1262016. View Article : Google Scholar : PubMed/NCBI
30	Hsu YT, Wolter KG and Youle RJ: Cytosol-to-membrane redistribution of Bax and Bcl-X(L) during apoptosis. Proc Natl Acad Sci USA. 94:pp. 3668–3672. 1997; View Article : Google Scholar : PubMed/NCBI
31	Boonstra J and Verkleij AJ: Regulation of enzyme activity in vivo is determined by its cellular localization. Adv Enzyme Regul. 44:61–73. 2004. View Article : Google Scholar : PubMed/NCBI
32	Zhao ZL, Li QF, Zheng YB, Chen LY, Shi SL and Jing GJ: The aberrant expressions of nuclear matrix proteins during the apoptosis of human osteosarcoma cells. Anat Rec (Hoboken). 293:813–820. 2010. View Article : Google Scholar : PubMed/NCBI
33	Bai Z, Ye Y, Liang B, Xu F, Zhang H, Zhang Y, Peng J, Shen D, Cui Z, Zhang Z and Wang S: Proteomics-based identification of a group of apoptosis-related proteins and biomarkers in gastric cancer. Int J Oncol. 38:375–383. 2011.PubMed/NCBI
34	Wyllie AH: Apoptosis (the 1992 Frank Rose Memorial Lecture). Br J Cancer. 67:205–208. 1993. View Article : Google Scholar : PubMed/NCBI
35	Bazhanova ED, Sukhanov DS and Teplyi DL: Pathways of apoptosis regulation in hepatocytes induced by first-line antitubercular drugs. Bull Exp Biol Med. 158:650–653. 2015. View Article : Google Scholar : PubMed/NCBI
36	Wu JS, Cheung WM, Tsai YS, Chen YT, Fong WH, Tsai HD, Chen YC, Liou JY, Shyue SK, Chen JJ, et al: Ligand-activated peroxisome proliferator-activated receptor-gamma protects against ischemic cerebral infarction and neuronal apoptosis by 14-3-3 epsilon upregulation. Circulation. 119:1124–1134. 2009. View Article : Google Scholar : PubMed/NCBI
37	Wong RS: Apoptosis in cancer: From pathogenesis to treatment. J Exp Clin Cancer Res. 30:872011. View Article : Google Scholar : PubMed/NCBI
38	Won J, Kim DY, La M, Kim D, Meadows GG and Joe CO: Cleavage of 14-3-3 protein by caspase-3 facilitates bad interaction with Bcl-x(L) during apoptosis. J Biol Chem. 278:19347–19351. 2003. View Article : Google Scholar : PubMed/NCBI
39	Samuel T, Weber HO, Rauch P, Verdoodt B, Eppel JT, McShea A, Hermeking H and Funk JO: The G2/M regulator 14-3-3sigma prevents apoptosis through sequestration of Bax. J Biol Chem. 276:45201–45206. 2001. View Article : Google Scholar : PubMed/NCBI
40	Selvakumaran M, Lin HK, Miyashita T, Wang HG, Krajewski S, Reed JC, Hoffman B and Liebermann D: Immediate early up-regulation of bax expression by p53 but not TGF beta 1: A paradigm for distinct apoptotic pathways. Oncogene. 9:1791–1798. 1994.PubMed/NCBI
41	Meek DW: The p53 response to DNA damage. DNA Repair (Amst). 3:1049–1056. 2004. View Article : Google Scholar : PubMed/NCBI
42	Fridman JS and Lowe SW: Control of apoptosis by p53. Oncogene. 22:9030–9040. 2003. View Article : Google Scholar : PubMed/NCBI
43	Gbormittah FO, Haab BB, Partyka K, Garcia-Ott C, Hancapie M and Hancock WS: Characterization of glycoproteins in pancreatic cyst fluid using a high-performance multiple lectin affinity chromatography platform. J Proteome Res. 13:289–299. 2014. View Article : Google Scholar : PubMed/NCBI
44	Milde-Langosch K: The Fos family of transcription factors and their role in tumourigenesis. Eur J Cancer. 41:2449–2461. 2005. View Article : Google Scholar : PubMed/NCBI
45	Smeyne RJ, Vendrell M, Hayward M, Baker SJ, Miao GG, Schilling K, Robertson LM, Curran T and Morgan JI: Continuous c-fos expression precedes programmed cell death in vivo. Nature. 363:166–169. 1993. View Article : Google Scholar : PubMed/NCBI
46	Chen X, Shen J, Wang Y, Chen X, Yu S, Shi H and Huo K: Up-regulation of c-Fos associated with neuronal apoptosis following intracerebral hemorrhage. Cell Mol Neurobiol. 35:363–376. 2015. View Article : Google Scholar : PubMed/NCBI
47	Yu AY, Su QR, Wang L, Zhou J and Liu XH: Effects of citalopram on the expression of PCNA and C-fos and cell apoptosis in rat frontal cortical neurons after stress. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 30:439–442. 2014.(In Chinese). PubMed/NCBI
48	Morales D, Skoulakis EC and Acevedo SF: 14-3-3s are potential biomarkers for HIV-related neurodegeneration. J Neurovirol. 18:341–353. 2012. View Article : Google Scholar : PubMed/NCBI
49	Gelman BB and Nguyen TP: Synaptic proteins linked to HIV-1 infection and immunoproteasome induction: Proteomic analysis of human synaptosomes. J Neuroimmune Pharmacol. 5:92–102. 2010. View Article : Google Scholar : PubMed/NCBI
50	Toyo-oka K, Shionoya A, Gambello MJ, Cardoso C, Leventer R, Ward HL, Ayala R, Tsai LH, Dobyns W, Ledbetter D, et al: 14-3-3epsilon is important for neuronal migration by binding to NUDEL: A molecular explanation for Miller-Dieker syndrome. Nat Genet. 34:274–285. 2003. View Article : Google Scholar : PubMed/NCBI