@article{fb1d65de320c45f0baeca11b4fdee564,
title = "Midkine induces epithelial-mesenchymal transition through Notch2/Jak2-Stat3 signaling in human keratinocytes",
abstract = "Epithelial-to-mesenchymal transition (EMT) underlies cell plasticity and embryonic development and is frequently observed in advanced tumorigenesis. We demonstrated that midkine (MK), a retinoic acid-inducible heparin-binding mitogen, promotes EMT in immortalized HaCaT keratinocytes. We showed that MK binds to the Notch2 receptor in HaCaT keratinocytes. We further found that MK activates Notch2 signaling leading to protein/protein interactions between Hes1 and Jak2/Stat3 intermediates. We thus suggest that MK-induced cross talk of Notch2/Jak2/Stat3 signaling pathways can regulate cell plasticity and motility contributing to the EMT and later stages of tumorigenesis.",
keywords = "EMT, Epithelial-to-mesenchymal transition, Immortalized HaCaT keratinocytes, Jak2, Midkine, Notch2, Squamous cell carcinomas, Stat3, shRNA",
author = "Yiping Huang and Hoque, {Mohammad Obaidul} and Feng Wu and Barry Trink and David Sidransky and Ratovitski, {Edward A.}",
note = "Funding Information: grown on cover slips and fixed in 4% formaldehyde in PBS (pH 7.4) Yuriditsky, Dafna Kisselman, Oksana Kazan and Michael (Mickie) We also thank undergraduate students Anna Brechman, Eugene for 30 min at room temperature. Cells were permeabilized for 10 min Leibovic for their help at the various stages of this study. These in 1% Triton X-100 in PBS. After washing with PBS, cells were incu-studies were supported in part by RPG-TBE-98317 grant from the bated for 30 min at room temperature with the indicated primary American Cancer Society (E.R.). . antibodies diluted in 1% FBS in PBS. After three washes in PBS, cells were incubated for 30 min with secondary conjugates (Alexa References 350 (blue)-labeled goat anti-rabbit IgG, Alexa 546 (red) -labeled goat 2.Birchmeier C, Birchmeier W, Brand-Saberi B. Epithelial-mesenchymal transitions in cancer 1.Hay E. An overview of epithelial-mesenchymal transformation. Acta Anat 1995; 154:8-20. anti-rabbit IgG or Alexa 488 (green)-labeled goat anti-mouse IgG as progression. Acta Anat 1996; 156:217-26. indicated. All secondary conjugates were diluted 1:500 in 1% FBS Huber MA, Kraut N, Beug H. Molecular requirements for epithelial-mesenchymal transi- in PBS. Cover slips were washed three times in PBS and mounted Leong KG, Karsan A. Recent insights into the role of Notch signaling in tumorigenesis. tion during tumor progression. Curr Opin Cell Biol 2005; 17:548-58. with ProLong antifading agent (Molecular Probes Inc.,). Confocal Blood 2006; 107:2223-33. microscopy was performed at the JHU Core Facility. Nuclei were Timmerman LA, Grego Bessa J, Raya A, Bertr{\'a}n E, P{\'e}rez Pomares JM, D{\'i}ez J, Aranda S, stained with Hoechst dye. mesenchymal transitioTn d uDringI ScarTdiaRc dIevBeloUpmTentE and oncogenic transformation. Genes Palomo S, McCormick F, Izpis{\'u}a Belmonte JC, de la Pompa JL. Notch promotes epithelial- Gene expression profiling. HaCaT cells were treated with physi-Dev 2004; 18:99-115. ologic concentrations of MK or control medium.26Total RNA was 6. ZavadilJ,CermakL,Soto-NievesN,B{\"o}ttingerEP.IntegrationofTGFβ/Smad and Jagged1/ isolated with RNA Easy purification kit (Qiagen) and after the 7. Fischer A, Schumacher N, Maier M, Sendtner M, Gessler M. The Notch target genes Hey1 Notch signalling in epithelial-to-mesenchymal transition. EMBO J 2004; 23:1155-65. biotinylation of cRNA probes (15 μg/chip) was hybridized with the and Hey2 are required for embryonic vascular development. Genes Dev 2004; 18:901-11. Affymetrix HU133A human gene chip array (22,283 oligonucle-. DO NO8.Zavadil J, Bitzer M, Liang D, Yang YC, Massimi A, Kneitz S, Piek E, Bottinger E. Genetic otide sequences) for 16 h at 45°C. Hybridized chips were incubated Acad Sci USA 2001; 98:6686-91.programs of epithelial cell plasticity directed by transforming growth factor-beta. Proc Natl the blots with a streptavidin-phycoerythrin conjugate followed by a 9. Nickoloff BJ, Qin JZ, Chaturvedi V, Denning MF, Bonish B, Miele L. Jagged-1 mediated labeling with an anti-streptavidin goat biotinylated antibody (Vector activation of Notch signaling induces complete maturation of human keratinocytes through Laboratories) and stain again with the streptavidin-phycoerythrin Mitsiadis TA, Lardelli M, Lendahl U, Thesleff I. Expression of Notch 1, 2 and 3 is regulated NFκB and PPARγ. Cell Death Differ 2002; 9:842-55. conjugate. Chips were then scanned using a Hewlett Packard scanner by epithelial-mesenchymal interactions and retinoic acid in the developing mouse tooth and by a photomultiplier tube through a 570 nm long pass filter. Digitized associated with determination of ameloblast cell fate. J Cell Biol 1995; 130:407-18. image data were processed using the Genechip software (Affymetrix, differentiation factor, is regulated by retinoic acid and epithelial-mesenchymal interactions Mitsiadis T, Muramatsu T, Muramatsu H, Thesleff I. Midkine, a heparin-binding growth/ Version 3.1). Scaled data obtained from scanner (experimental condi-in the developing mouse tooth, and affects cell proliferation and morphogenesis. J Cell Biol tions) were normalized to the data obtained from normal untreated 1995; 198:267-81. conditions, and specific genes that were changed in expression with Lelievre-Pegorier M. Midkine is involved in kidney development and in its regulation by Vilar J, Lalou C, Duong VH, Charrin S, Hardouin S, Raulais D, Merlet-Benichou C, more than a 2-fold change were tabulated (NOMAD). We used three retinoids. J Am Soc Nephrol 2002; 13:668-76. biological replicas for each of our samples and considered genes for Matsuura O, Kadomatsu K, Takei Y, Uchimura K, Mimura S, Watanabe K, Muramatsu T. further analysis that showed common pattern of fold changes in 2002; 2:109-15.Midkine expression is associated with postnatal development of the lungs. Cell Struct Funct each of the three individual experiments. The statistical analysis of Morrisey EE, Savani RC. Midkine: a potential bridge between glucocorticoid and retinoid microarrays was used to identify significant gene targets validated by effects on lung vascular development. Am J Respir Cell Mol Biol 2003; 1:5-8. ANOVA (corrected), PCA, and Hierarchical clustering analysis. induces the transformation of NIH3T3 cells. Br J Cancer 1997; 3:354-9.15.Kadomatsu K, Hagihara M, Akhter S, Fan Q, Muramatsu H, Muramatsu T. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.",
year = "2008",
month = jun,
day = "1",
doi = "10.4161/cc.7.11.5952",
language = "English (US)",
volume = "7",
pages = "1613--1622",
journal = "Cell Cycle",
issn = "1538-4101",
publisher = "Landes Bioscience",
number = "11",
}