Abstract
Aim:
Protein L-isoaspartyl O-methyltransferase (PIMT) regulates cell adhesion in various cancer cell lines through activation of integrin αv and the PI3K pathway. The epithelial mesenchymal transition (EMT) enables epithelial cells to acquire the characteristics of mesenchymal cells, and to allow them to migrate for metastasis. Here, we examined the relationship between PIMT and EMT with attached or detached MDA-MB 231 cells.
Methods:
Human breast cancer cell line MDA-MB-231 cells were maintained in a suspension on poly-HEMA in the presence or absence of PIMT siRNA or ERK inhibitor PD98059. The mRNAs and proteins were analyzed using RT-PCR and immunoblotting, respectively.
Results:
During cellular incubation under detached conditions, PIMT, integrin αv and EMT proteins, such as Snail, Slug and matrix metalloproteinase 2 (MMP-2), were significantly increased in correlation with the phosphorylation of ERK1/2. The ERK inhibitor PD98059 (25 μmol/L) strongly suppressed the expression of the proteins and PIMT. Interestingly, PIMT siRNA blocked the phosphorylation of ERK and the expression of the EMT proteins. Additionally, PIMT and ERK phosphorylation were both co-activated by treatment with TGF-β (10 ng/mL) and TNF-α (10 ng/mL).
Conclusion:
A tight cross-regulation exists between ERK and PIMT in regards to their activation and expression during the EMT.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Creighton CJ, Chang JC, Rosen JM . Epithelial-mesenchymal transition (EMT) in tumor-initiating cells and its clinical implications in breast cancer. J Mammary Gland Biol Neoplasia 2010; 15: 253–60.
Yang J, Weinberg RA . Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Dev Cell 2008; 14: 818–29.
Do Kwon Y, Bin Noh H, Ahn HJ, Lee WJ, Kwack K . The molecular signature of in vitro senescence in human mesenchymal stem cells. Genes Genom 2010; 32: 87–93.
Gotzmann J, Mikula M, Eger A, Schulte-Hermann R, Foisner R, Beug H, et al. Molecular aspects of epithelial cell plasticity: implications for local tumor invasion and metastasis. Mutat Res 2004; 566: 9–20.
Smalley KS, Brafford PA, Herlyn M . Selective evolutionary pressure from the tissue microenvironment drives tumor progression. Semin Cancer Biol 2005; 15: 451–9.
Shimizu T, Matsuoka Y, Shirasawa T . Biological significance of isoaspartate and its repair system. Biol Pharm Bull 2005; 28: 1590–6.
Reissner KJ, Aswad DW . Deamidation and isoaspartate formation in proteins: unwanted alterations or surreptitious signals? Cell Mol Life Sci 2003; 60: 1281–95.
Lanthier J, Desrosiers RR . Protein L-isoaspartyl methyltransferase repairs abnormal aspartyl residues accumulated in vivo in type-I collagen and restores cell migration. Exp Cell Res 2004; 293: 96–105.
Lanthier J, Desrosiers RR . Regulation of protein L-isoaspartyl methyltransferase by cell-matrix interactions: involvement of integrin alphavbeta3, PI3-kinase, and the proteasome. Biochem Cell Biol 2006; 84: 684–94.
Johnson-Holiday C, Singh R, Johnson E, Singh S, Stockard CR, Grizzle WE, et al. CCL25 mediates migration, invasion and matrix metalloproteinase expression by breast cancer cells in a CCR9-dependent fashion. Int J Oncol 2011; 38: 1279–85.
Garrido T, Riese HH, Aracil M, Perez-Aranda A . Endothelial cell differentiation into capillary-like structures in response to tumour cell conditioned medium: a modified chemotaxis chamber assay. Br J Cancer 1995; 71: 770–5.
Hunakova L, Sedlakova O, Cholujova D, Gronesova P, Duraj J, Sedlak J . Modulation of markers associated with aggressive phenotype in MDA-MB-231 breast carcinoma cells by sulforaphane. Neoplasma 2009; 56: 548–56.
Cho JY, Fox DA, Horejsi V, Sagawa K, Skubitz KM, Katz DR, et al. The functional interactions between CD98, beta1-integrins, and CD147 in the induction of U937 homotypic aggregation. Blood 2001; 98: 374–82.
Rhee DK, Kim EH, Lee MJ, Kim IH, Pyo S, Choi KT . Anti-apoptotic effects of red ginseng on oxidative stress induced by hydrogen peroxide in SK-N-SH cells. J Ginseng Res 2010; 34: 138–44.
Lee YH, Hong JK, Kim JS, Kim JA, Lee SI, Lim MH, et al. Identification and characterization of SHI family genes from Brassica rapa L ssp pekinensis. Genes Genom 2010; 32: 309–17.
Xing P, Li JG, Jin F, Zhao TT, Liu Q, Dong HT, et al. Fascin, an actin-bundling protein, promotes breast cancer progression in vitro. Cell Biochem Funct 2011; 29: 303–10.
Park JK, Do Kim H, Ha SE, Kang JR . Effect of Korean red ginseng extract on cell death responses in peroxynitrite-treated keratinocytes. J Ginseng Res 2010; 34: 205–11.
Tsuchiyama J, Imajo K, Yoshino T, Nanba N, Toyota A, Yoshida C, et al. High-dose chemotherapy and autologous peripheral blood stem cell transplantation for treatment of unspecified peripheral T-cell lymphoma presented with hepatosplenomegaly and hypercytokinemia syndrome: report of three cases. Ann Hematol 2002; 81: 588–92.
Chun YJ, Shim JH, Choi CS, Lee EC, Kim MY . Tamoxifen suppresses clusterin level through Akt inactivation and proteasome degradation in human prostate cancer cells. Biomol Ther 2009; 17: 25–31.
Chun YJ, Lee HM, Surh BY . Roles of neutral sphingomyelinase 1 on CD95-mediated apoptosis in human Jurkat T lymphocytes. Biomol Ther 2010; 18: 262–70.
Gottschall PE, Yu X . Cytokines regulate gelatinase A and B (matrix metalloproteinase 2 and 9) activity in cultured rat astrocytes. J Neurochem 1995; 64: 1513–20.
Moon A, Kim JS . Fibronectin induces Pro-MMP-2 activation and enhances invasion in H-Ras-transformed human breast epithelial cells. Biomol Ther 2009; 17: 288–92.
Nobes CD, Hall A . Rho GTPases control polarity, protrusion, and adhesion during cell movement. J Cell Biol 1999; 144: 1235–44.
Kang TJ, Lee GS, Choi JY, Choi YJ, Yim DS, Cheong JH . The wound healing effect of Hydnocarpi Semen extract on ulcer in diabetic mice. Biomol Ther 2010; 18: 329–35.
Haraguchi M, Okubo T, Miyashita Y, Miyamoto Y, Hayashi M, Crotti TN, et al. Snail regulates cell-matrix adhesion by regulation of the expression of integrins and basement membrane proteins. J Biol Chem 2008; 283: 23514–23.
Larriba MJ, Martin-Villar E, Garcia JM, Pereira F, Pena C, de Herreros AG, et al. Snail2 cooperates with Snail1 in the repression of vitamin D receptor in colon cancer. Carcinogenesis 2009; 30: 1459–68.
Lamarre M, Desrosiers RR . Up-regulation of protein L-isoaspartyl methyltransferase expression by lithium is mediated by glycogen synthase kinase-3 inactivation and beta-catenin stabilization. Neuropharmacology 2008; 55: 669–76.
Arima Y, Inoue Y, Shibata T, Hayashi H, Nagano O, Saya H, et al. Rb depletion results in deregulation of E-cadherin and induction of cellular phenotypic changes that are characteristic of the epithelial-to-mesenchymal transition. Cancer Res 2008; 68: 5104–12.
Kim JB, Yu JH, Ko E, Lee KW, Song AK, Park SY, et al. The alkaloid berberine inhibits the growth of Anoikis-resistant MCF-7 and MDA-MB-231 breast cancer cell lines by inducing cell cycle arrest. Phytomedicine 2010; 17: 436–40.
Savagner P . The epithelial-mesenchymal transition (EMT) phenomenon. Ann Oncol 2010; 21: vii89–vii92.
Thiery JP . Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2002; 2: 442–54.
Manes T, Zheng DQ, Tognin S, Woodard AS, Marchisio PC, Languino LR . Alpha(v)beta3 integrin expression up-regulates cdc2, which modulates cell migration. J Cell Biol 2003; 161: 817–26.
Chou TY, Chen WC, Lee AC, Hung SM, Shih NY, Chen MY . Clusterin silencing in human lung adenocarcinoma cells induces a mesenchymal-to-epithelial transition through modulating the ERK/Slug pathway. Cell Signal 2009; 21: 704–11.
Kuo PL, Chen YH, Chen TC, Shen KH, Hsu YL . CXCL5/ENA78 increased cell migration and epithelial-to-mesenchymal transition of hormone-independent prostate cancer by early growth response-1/Snail signaling pathway. J Cell Physiol 2010; 226: 1224–31.
Gollob JA, Wilhelm S, Carter C, Kelley SL . Role of Raf kinase in cancer: therapeutic potential of targeting the Raf/MEK/ERK signal transduction pathway. Semin Oncol 2006; 33: 392–406.
Cho JY, Skubitz KM, Katz DR, Chain BM . CD98-dependent homotypic aggregation is associated with translocation of protein kinase Cdelta and activation of mitogen-activated protein kinases. Exp Cell Res 2003; 286: 1–11.
Stahle M, Veit C, Bachfischer U, Schierling K, Skripczynski B, Hall A, et al. Mechanisms in LPA-induced tumor cell migration: critical role of phosphorylated ERK. J Cell Sci 2003; 116: 3835–46.
Gal A, Sjoblom T, Fedorova L, Imreh S, Beug H, Moustakas A . Sustained TGF beta exposure suppresses Smad and non-Smad signalling in mammary epithelial cells, leading to EMT and inhibition of growth arrest and apoptosis. Oncogene 2008; 27: 1218–30.
Huebscher KJ, Lee J, Rovelli G, Ludin B, Matus A, Stauffer D, et al. Protein isoaspartyl methyltransferase protects from Bax-induced apoptosis. Gene 1999; 240: 333–41.
Furuchi T, Sakurako K, Katane M, Sekine M, Homma H . The role of protein L-isoaspartyl/D-aspartyl O-methyltransferase (PIMT) in intracellular signal transduction. Chem Biodivers 2010; 7: 1337–48.
Cournoyer P, Desrosiers RR . Valproic acid enhances protein L-isoaspartyl methyltransferase expression by stimulating extracellular signal-regulated kinase signaling pathway. Neuropharmacology 2009; 56: 839–48.
Alison MR, Lim SM, Nicholson LJ . Cancer stem cells: problems for therapy? J Pathol 2011; 223: 147–61.
Kong D, Banerjee S, Ahmad A, Li Y, Wang Z, Sethi S, et al. Epithelial to mesenchymal transition is mechanistically linked with stem cell signatures in prostate cancer cells. PLoS One 2010; 5: e12445.
Kondo M, Wagers AJ, Manz MG, Prohaska SS, Scherer DC, Beilhack GF, et al. Biology of hematopoietic stem cells and progenitors: implications for clinical application. Annu Rev Immunol 2003; 21: 759–806.
Bhat-Nakshatri P, Appaiah H, Ballas C, Pick-Franke P, Goulet R Jr, Badve S, et al. SLUG/SNAI2 and tumor necrosis factor generate breast cells with CD44+/CD24− phenotype. BMC Cancer 2010; 10: 411.
Acknowledgements
This research was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Ryu, J., Song, J., Heo, J. et al. Cross-regulation between protein L-isoaspartyl O-methyltransferase and ERK in epithelial mesenchymal transition of MDA-MB-231 cells. Acta Pharmacol Sin 32, 1165–1172 (2011). https://doi.org/10.1038/aps.2011.94
Received:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/aps.2011.94
Keywords
This article is cited by
-
The enzyme l-isoaspartyl (d-aspartyl) methyltransferase is required for VEGF-dependent endothelial cell migration and tubulogenesis
Molecular and Cellular Biochemistry (2016)
