Ukr.Biochem.J. 2018; Volume 90, Issue 3, May-Jun, pp. 94-100

doi: https://doi.org/10.15407/ubj90.03.094

Adaptor protein Ruk/CIN85 modulates resistance to doxorubicin of murine 4T1 breast cancer cells

I. R. Horak, D. S. Gerashchenko, L. B. Drobot

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: iryna.horak@gmail.com

The acquisition of chemoresistance in the course of tumor progression includes activation of membrane ABC transporters, detoxification enzymes, cell cycle deceleration and activation of specific signaling pathways such as Akt/mTOR, MAPK, NF-κB. Adaptor proteins play an essential role in the assembly of supramolecular signaling complexes, maintaining and directing the intracellular signaling. One of such proteins, called Ruk/CIN85, is strongly associated with malignant transformation and metastasis. In present study we investigated the Ruk/CIN85 effect of up/down-regulation on the transforming potential and doxorubicin resistance of highly aggressive mouse breast adenocarcinoma 4T1 cells. It was demonstrated that 4T1 cells overexpressing Ruk/CIN85 possessed increased resistance to doxorubicin (in the range of concentrations 0.1–10.0 µM) while knockdown cells were the most sensitive. Also, high levels of Ruk/CIN85 in 4T1 cells positively correlated with their ability to form colonies in semi-solid agar. Ruk/CIN85-overexpressing cells formed four times more colonies in comparison with Ruk/CIN85 nockdown cells, the growth of which revealed higher resistance to doxorubicin action.

Keywords: , , ,


References:

  1. Jackson SE, Chester JD. Personalised cancer medicine. Int J Cancer. 2015 Jul 15;137(2):262-6.  PubMed, CrossRef
  2. Vanneman M, Dranoff G. Combining immunotherapy and targeted therapies in cancer treatment. Nat Rev Cancer. 2012 Mar 22;12(4):237-51. PubMed, PubMedCentral, CrossRef
  3. Wang T, Shigdar S, Gantier MP, Hou Y, Wang L, Li Y, Shamaileh HA, Yin W, Zhou SF, Zhao X, Duan W. Cancer stem cell targeted therapy: progress amid controversies. Oncotarget. 2015 Dec 29;6(42):44191-206. PubMed, PubMedCentral, CrossRef
  4. Monroig Pdel C, Chen L, Zhang S, Calin GA. Small molecule compounds targeting miRNAs for cancer therapy. Adv Drug Deliv Rev. 2015 Jan;81:104-16. PubMed, PubMedCentral, CrossRef
  5. Mitra AK, Agrahari V, Mandal A, Cholkar K, Natarajan C, Shah S, Joseph M, Trinh HM, Vaishya R, Yang X, Hao Y, Khurana V, Pal D. Novel delivery approaches for cancer therapeutics. J Control Release. 2015 Dec 10;219:248-268. PubMed, PubMedCentral, CrossRef
  6. Fisher R, Pusztai L, Swanton C. Cancer heterogeneity: implications for targeted therapeutics. Br J Cancer. 2013 Feb 19;108(3):479-85. PubMed, PubMedCentral, CrossRef
  7. Garraway LA, Jänne PA. Circumventing cancer drug resistance in the era of personalized medicine. Cancer Discov. 2012 Mar;2(3):214-26. PubMed, CrossRef
  8. Westover D, Li F. New trends for overcoming ABCG2/BCRP-mediated resistance to cancer therapies. J Exp Clin Cancer Res. 2015 Dec 30;34:159. PubMed, PubMedCentral, CrossRef
  9. Du B, Shim JS. Targeting Epithelial-Mesenchymal Transition (EMT) to Overcome Drug Resistance in Cancer. Molecules. 2016 Jul 22;21(7). pii: E965.
    PubMed, CrossRef
  10. Dikic I. CIN85/CMS family of adaptor molecules. FEBS Lett. 2002 Oct 2;529(1):110-5. PubMed, CrossRef
  11. Buchman VL, Luke C, Borthwick EB, Gout I, Ninkina N. Organization of the mouse Ruk locus and expression of isoforms in mouse tissues. Gene. 2002 Jul 24;295(1):13-17. PubMed, CrossRef
  12. Finniss S, Movsisyan A, Billecke C, Schmidt M, Randazzo L, Chen B, Bögler O. Studying protein isoforms of the adaptor SETA/CIN85/Ruk with monoclonal antibodies. Biochem Biophys Res Commun. 2004 Dec 3;325(1):174-82. PubMed, CrossRef
  13. Havrylov S, Ichioka F, Powell K, Borthwick EB, Baranska J, Maki M, Buchman VL. Adaptor protein Ruk/CIN85 is associated with a subset of COPI-coated membranes of the Golgi complex. Traffic. 2008 May;9(5):798-812.  PubMed, CrossRef
  14. Schroeder B, Srivatsan S, Shaw A, Billadeau D, McNiven MA. CIN85 phosphorylation is essential for EGFR ubiquitination and sorting into multivesicular bodies. Mol Biol Cell. 2012 Sep;23(18):3602-11. PubMed, PubMedCentral, CrossRef
  15. Ahmad G, Mohapatra BC, Schulte NA, Nadeau SA, Luan H, Zutshi N, Tom E, Ortega-Cava C, Tu C, Sanada M, Ogawa S, Toews ML, Band V, Band H. Cbl-family ubiquitin ligases and their recruitment of CIN85 are largely dispensable for epidermal growth factor receptor endocytosis. Int J Biochem Cell Biol. 2014 Dec;57:123-34.  PubMed, PubMedCentral, CrossRef
  16. Narita T, Nishimura T, Yoshizaki K, Taniyama T. CIN85 associates with TNF receptor 1 via Src and modulates TNF-alpha-induced apoptosis. Exp Cell Res. 2005 Mar 10;304(1):256-64. PubMed, CrossRef
  17. Havrylov S, Redowicz MJ, Buchman VL. Emerging roles of Ruk/CIN85 in vesicle-mediated transport, adhesion, migration and malignancy. Traffic. 2010 Jun;11(6):721-31.  PubMed, CrossRef
  18. Bai SW, Herrera-Abreu MT, Rohn JL, Racine V, Tajadura V, Suryavanshi N, Bechtel S, Wiemann S, Baum B, Ridley AJ. Identification and characterization of a set of conserved and new regulators of cytoskeletal organization, cell morphology and migration. BMC Biol. 2011 Aug 11;9:54. PubMed, PubMedCentral, CrossRef
  19. Yasin HWR, van Rensburg SH, Feiler CE, Johnson RI. The adaptor protein Cindr regulates JNK activity to maintain epithelial sheet integrity. Dev Biol. 2016 Feb 15;410(2):135-149. PubMed, CrossRef
  20. Cascio S, Finn OJ.  Complex of MUC1, CIN85 and Cbl in Colon Cancer Progression and Metastasis. Cancers (Basel). 2015 Feb 10;7(1):342-52. PubMed, PubMedCentral, CrossRef
  21. Ma Y, Ye F, Xie X, Zhou C, Lu W. Significance of PTPRZ1 and CIN85 expression in cervical carcinoma. Arch Gynecol Obstet. 2011 Sep;284(3):699-704. PubMed, CrossRef
  22. 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 Oct;33(10):1976-84. PubMed, CrossRef
  23. Gout I, Middleton G, Adu J, Ninkina NN, Drobot LB, Filonenko V, Matsuka G, Davies AM, Waterfield M, Buchman VL. Negative regulation of PI 3-kinase by Ruk, a novel adaptor protein. EMBO J. 2000 Aug 1;19(15):4015-25. PubMed, PubMedCentral, CrossRef
  24. 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 responces of tumor cells. Biotechnol Acta. 2013; 6(4): 182-189.  CrossRef
  25. Borowicz S, Van Scoyk M, Avasarala S, Karuppusamy Rathinam MK, Tauler J, Bikkavilli RK, Winn RA. The soft agar colony formation assay. J Vis Exp. 2014 Oct 27;(92):e51998.  PubMed, PubMedCentral, CrossRef
  26. Cojoc M, Mäbert K, Muders MH, Dubrovska A. A role for cancer stem cells in therapy resistance: cellular and molecular mechanisms. Semin Cancer Biol. 2015 Apr;31:16-27. PubMed, CrossRef
  27. Bögler O, Furnari FB, Kindler-Roehrborn A, Sykes VW, Yung R, Huang HJ, Cavenee WK. SETA: a novel SH3 domain-containing adapter molecule associated with malignancy in astrocytes. Neuro Oncol. 2000 Jan;2(1):6-15. PubMed, PubMedCentral, CrossRef
  28. Cascio S, Farkas AM, Hughey RP, Finn OJ. Altered glycosylation of MUC1 influences its association with CIN85: the role of this novel complex in cancer cell invasion and migration. Oncotarget. 2013 Oct;4(10):1686-97. PubMed, PubMedCentral, CrossRef
  29. Sokolosky ML, Stadelman KM, Chappell WH, Abrams SL, Martelli AM, Stivala F, Libra M, Nicoletti F, Drobot LB, Franklin RA, Steelman LS, McCubrey JA. Involvement of Akt-1 and mTOR in sensitivity of breast cancer to targeted therapy. Oncotarget. 2011 Jul;2(7):538-50. PubMed, PubMedCentral, CrossRef
  30. Niero EL, Rocha-Sales B, Lauand C, Cortez BA, de Souza MM, Rezende-Teixeira P, Urabayashi MS, Martens AA, Neves JH, Machado-Santelli GM. The multiple facets of drug resistance: one history, different approaches. J Exp Clin Cancer Res. 2014 Apr 28;33:37. PubMed, PubMedCentral, CrossRef
  31. McCubrey JA, Abrams SL, Fitzgerald TL, Cocco L, Martelli AM, Montalto G, Cervello M, Scalisi A, Candido S, Libra M, Steelman LS. Roles of signaling pathways in drug resistance, cancer initiating cells and cancer progression and metastasis. Adv Biol Regul. 2015 Jan;57:75-101.  PubMed, CrossRef
  32. Pasichnyk GV, Povorozniuk OO, Horak IR, Gerashchenko DS, Ponomarenko OV, Samoylenko AA, Byts NV, Drobot LB. Overexpression of adaptor protein Rukl/CIN85 in human breast adenocarcinoma cell line MCF-7 is accompanied by increased chemoresistance. Rep Nat Acad Sci Ukraine. 2013; 12: 149–156. (In Ukrainian).
  33. AbuHammad S, Zihlif M. Gene expression alterations in doxorubicin resistant MCF7 breast cancer cell line. Genomics. 2013 Apr;101(4):213-20. PubMed, CrossRef

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