Biopolym. Cell. 2016; 32(1):21-25.
Structure and Function of Biopolymers
Apocynin attenuates motility and induces transition from sustained to transient EGF-dependent Akt activation in MCF-7 cells that overexpress adaptor protein Ruk/CIN85
1Bazalii A. V., 1Drobot L. B., 1Komisarenko S. V.
  1. Palladin Institute of Biochemistry, NAS of Ukraine
    9, Leontovycha Str., Kyiv, Ukraine, 01601


Aim. To study a possible involvement of NADPH oxidases in the control of cell motility and Akt signaling in the human breast adenocarcinoma MCF-7 cells that stably overexpress the full-length form of adaptor protein Ruk/CIN85. Methods. Cell motility was studied by a transwell migration assay. The dynamics of EGF-induced Akt activation was investigated by Western blot analysis. Results. It has been shown that apocynin, an inhibitor of the assembly of plasma membrane NADPH oxidases, substantially attenuates the motility of Ruk/CIN85 overexpressing MCF-7 cells (subclone G10) in comparison with control cells. In addition, apocynin induced the transition from sustained to transient EGF-dependent Akt activation in G10 cells and did not influence transient Akt activation in control cells. Conclusions. The data obtained can suggest that ROS produced by NADPH oxidases are signaling components, upstream to Akt kinase, that mediate the increased migratory potential of Ruk/CIN85 overexpressing MCF-7 cells.
Keywords: NADPH oxidases, apocynin, adaptor protein Ruk/CIN85, motility, EGF, Akt signaling


[1] Meitzler JL, Antony S, Wu Y, Juhasz A, Liu H, Jiang G, Lu J, Roy K, Doroshow JH. NADPH oxidases: a perspective on reactive oxygen species production in tumor biology. Antioxid Redox Signal. 2014;20(17):2873-89.
[2] Díaz B, Courtneidge SA. Redox signaling at invasive microdomains in cancer cells. Free Radic Biol Med. 2012;52(2):247-56.
[3] Roy K, Wu Y, Meitzler JL, Juhasz A, Liu H, Jiang G, Lu J, Antony S, Doroshow JH. NADPH oxidases and cancer. Clin Sci (Lond). 2015;128(12):863-75.
[4] Lambeth JD. NOX enzymes and the biology of reactive oxygen. Nat Rev Immunol. 2004;4(3):181-9.
[5] Katsuyama M. NOX/NADPH oxidase, the superoxide-generating enzyme: its transcriptional regulation and physiological roles. J Pharmacol Sci. 2010;114(2):134-46.
[6] Finkel T. Reactive oxygen species and signal transduction. IUBMB Life. 2001;52(1-2):3-6.
[7] Drobot LB, Samoylenko AA, Vorotnikov AV, Tyurin-Kuzmin PA, Bazalii AV, Kietzmann T, Tkachuk VA, Komisarenko SV. Reactive oxygen species in signal transduction. Ukr Biokhim Zh. 2013;85(6):209–17.
[8] Pawson T. Dynamic control of signaling by modular adaptor proteins. Curr Opin Cell Biol. 2007;19(2):112-6.
[9] Levchenko A, Bruck J, Sternberg PW. Scaffold proteins may biphasically affect the levels of mitogen-activated protein kinase signaling and reduce its threshold properties. Proc Natl Acad Sci U S A. 2000;97(11):5818-23.
[10] Havrylov S, Rzhepetskyy Y, Malinowska A, Drobot L, Redowicz MJ. Proteins recruited by SH3 domains of Ruk/CIN85 adaptor identified by LC-MS/MS. Proteome Sci. 2009;7:21.
[11] Bazalii AV, Samoylenko AA, Petukhov DM, Rynditch AV, Redowicz M-J, Drobot LB. Interaction between adaptor proteins Ruk. CIN85 and Tks4 in normal and tumor cells of different tissue origins. Biopolym Cell. 2014; 30(1): 37–41.
[12] Gianni D, Diaz B, Taulet N, Fowler B, Courtneidge SA, Bokoch GM. Novel p47(phox)-related organizers regulate localized NADPH oxidase 1 (Nox1) activity. Sci Signal. 2009;2(88):ra54.
[13] Gianni D, DerMardirossian C, Bokoch GM. Direct interaction between Tks proteins and the N-terminal proline-rich region (PRR) of NoxA1 mediates Nox1-dependent ROS generation. Eur J Cell Biol. 2011;90(2-3):164-71.
[14] Samoylenko AA, Byts NV, Pasichnyk GV, Kozlova NV, Bazalii AV, Gerashchenko DS, Shandrenko SG, Vorotnikov AV, Kietzmann T, Komisarenko SV, Drobot LB. Recombinant lentivirus-mediated silencing of adaptor protein Ruk. CIN85 expression influences biological responses of tumor cells. Biotechnologia Acta. 2013; 6(4): 182–9.
[15] Bazalii AV, Horak IR, Pasichnyk GV, Komisarenko SV, Drobot LB. Transcriptional regulation of NOX genes expression in human breast adenocarcinoma MCF-7 cells is modulated by adaptor protein Ruk. CIN85. Ukr Biochem J. 2016; 88(1): 119–25.
[16] Samoylenko A, Vynnytska-Myronovska B, Byts N, Kozlova N, Basaraba O, Pasichnyk G, Palyvoda K, Bobak Y, Barska M, Mayevska O, Rzhepetsky Y, Shuvayeva H, Lyzogubov V, Usenko V, Savran V, Volodko N, Buchman V, Kietzmann T, Drobot L. Increased levels of the HER1 adaptor protein Rukl/CIN85 contribute to breast cancer malignancy. Carcinogenesis. 2012;33(10):1976-84.
[17] Bazalii AV, Vorotnikov AV, Tiurin-Kuzmin PA, Tkachuk VA, Komisarenko SV, Drobot LB. Recombinant fluorescent sensor of hydrogen peroxide fused with adaptor protein Ruk. CIN85: desining of expression vector and functional characterization. Biotechnologia Acta. 2015; 8(5): 19–26.
[18] Ushio-Fukai M. Compartmentalization of redox signaling through NADPH oxidase-derived ROS. Antioxid Redox Signal. 2009;11(6):1289-99.
[19] Dikic I. CIN85/CMS family of adaptor molecules. FEBS Lett. 2002;529(1):110-5.
[20] Havrylov S, Redowicz MJ, Buchman VL. Emerging roles of Ruk/CIN85 in vesicle-mediated transport, adhesion, migration and malignancy. Traffic. 2010;11(6):721-31.
[21] Stefanska J, Pawliczak R. Apocynin: molecular aptitudes. Mediators Inflamm. 2008;2008:106507.
[22] McAuliffe PF, Meric-Bernstam F, Mills GB, Gonzalez-Angulo AM. Deciphering the role of PI3K/Akt/mTOR pathway in breast cancer biology and pathogenesis. Clin Breast Cancer. 2010;10 Suppl 3:S59-65.