, Volume 24, Issue 11–12, pp 958–971 | Cite as

Integrin-EGFR interaction regulates anoikis resistance in colon cancer cells

  • Deblina Guha
  • Taniya Saha
  • Sayantan Bose
  • Sreeparna Chakraborty
  • Subhanki Dhar
  • Poulami Khan
  • Arghya Adhikary
  • Tanya Das
  • Gaurisankar SaEmail author


Anoikis resistance is an essential property of cancer cells that allow the extra-cellular matrix-detached cells to survive in a suspended state in body fluid in order to metastasize and invade to distant organs. It is known that integrins play an important role in anoikis resistance, but detailed mechanisms are not well understood. Here we report that highly metastatic colon cancer cells showed a higher degree of anoikis resistance than the normal intestinal epithelial cells. These anoikis-resistant cancer cells express high-levels of integrin-α2, β1, and activated EGFR in the anchorage-independent state than the anchorage-dependent state. In contrast, normal intestinal epithelial cells failed to elevate these proteins. Interestingly, a higher co-association of EGFR with integrin-α2β1/-α5β1 was observed on the surface of anoikis-resistant cells. Thus, in the absence of extra-cellular matrix, integrins in association with EGFR activates downstream effectors ERK and AKT and suppress Caspase-3 activation to induce anoikis resistance as was confirmed from the gene-ablation and pharmacological inhibitor studies. Interestingly, these anoikis-resistant cancer cells express high-level of cancer stem cell signatures (CD24, CD44, CD133, EpCAM) and pluripotent stem cell markers (OCT-4, SOX-2, Nanog) as well as drug-resistant pumps (ABCG2, MDR1, MRP1). Altogether, our findings unravel the interplay between integrin-α2β1/-α5β1 and EGFR in anoikis resistance and suggest that the resistant cells are cancer initiating or cancer stem cells, which may serve as a promising target to combat metastasis of cancer.


Integrin EGFR Anoikis resistance Poly-HEMA HCT116 



We acknowledge Dr. Susanta Roychoudhury for providing the HCT116 cell line. Authors are thankful to Mr. R. Dutta, Mr. A. Poddar, Mrs. S. Das, and Mr. S. Chakrabarty, for their technical help. Our work was supported by the grants from the Department of Science & Technology and University Grants Commission, Govt. of India.

Author contributions

DG formulated the project, designed and executed most of the experiments, analyzed related results and wrote the manuscript; TS performed the immunostaining experiments and analyzed the results; SB assisted in cell-culture maintenance; SC analyzed experimental results; SD performed cell viability assays and Western blot experiments; PK analyzed experimental results; AA helped conceptually and gave critical suggestions; TD designed the experiments and edited the manuscript; GS conceptualized the project, designed the experiments, edited the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and animals

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    Day ML, Foster RG, Day KC, Zhao X, Humphrey P, Swanson P et al (1997) Cell anchorage regulates apoptosis through the retinoblastoma tumor suppressor/E2F pathway. J Biol Chem 272:8125–8128CrossRefPubMedGoogle Scholar
  2. 2.
    Ranger AM, Zha J, Harada H, Datta SR, Danial NN, Gilmore, AP et al (2003) Bad-deficient mice develop diffuse large B cell lymphoma. Proc Natl Acad Sci USA. 100:9324–9329CrossRefPubMedGoogle Scholar
  3. 3.
    Dufour G, Demers MJ, Gagne D, Dydensborg AB, Teller IC, Bouchard V et al (2004) Human intestinal epithelial cell survival and anoikis: differentiation state distinct regulation and roles of protein kinase B/Akt isoforms. J Biol Chem 279:44113–44122CrossRefPubMedGoogle Scholar
  4. 4.
    Aplin AE, Howe AK, Juliano RL (1999) Cell adhesion molecules, signal transduction, and cell growth. Curr Opin Cell Biol 11:737–744CrossRefPubMedGoogle Scholar
  5. 5.
    Meredith JE Jr, Fazeli B, Schwartz MA (1993) The extracellular matrix as a cell survival factor. Mol Biol Cell 4:953–961CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Zhang Z, Vuori K, Reed JC, Ruoslahti E (1995) The alpha5 beta1 integrin supports the survival of cells on fibronectin and up-regulates Bcl-2 expression. Proc Natl Acad Sci USA 92:6161–6165CrossRefPubMedGoogle Scholar
  7. 7.
    O’Brien V, Frisch SM, Juliano RL (1996) Expression of the integrin alpha5 subunit in HT29 colon carcinoma cells suppresses apoptosis triggered by serum deprivation. Exp Cell Res 224:208–213CrossRefPubMedGoogle Scholar
  8. 8.
    Wary KK, Mainiero F, Isakoff SJ, Marcantonio EE, Giancotti FG (1996) The adaptor protein Shc couples a class of integrins to the control of cell cycle progression. Cell 87:733–743CrossRefPubMedGoogle Scholar
  9. 9.
    Marco RA, Díaz-Montero CM, Wygant JN, Kleinerman ES, McIntyre BW (2003) Alpha4 integrin increases anoikis of human osteosarcoma cells. J Cell Biochem 88:1038–1047CrossRefPubMedGoogle Scholar
  10. 10.
    Benoit YD, Larrivée JF, Groulx JF, Stankova J, Vachon PH, Beaulieu JF et al (2010) Integrin α8β1 confers anoikis susceptibility to human intestinal epithelial crypt cells. Biochem Biophys Res Commun 399(3):434–439CrossRefPubMedGoogle Scholar
  11. 11.
    Moro L, Venturino M, Bozzo C, Silengo L, Altruda F, Beguinot L et al (1998) Integrins induce activation of EGF receptor: role in MAP kinase induction and adhesion-dependent cell survival. EMBO J 17(22):6622–6632CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Schneller M, Vuori K, Ruoslahti E (1997) Alphavbeta3 integrin associates with activated insulin and PDGFbeta receptors and potentiates the biological activity of PDGF. EMBO J 16(18):5600–5607CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Riedl SJ, Salvesen GS (2007) The apoptosome: signalling platform of cell death. Nat Rev Mol Cell Biol 8(5):405–413CrossRefPubMedGoogle Scholar
  14. 14.
    Hehlgans S, Haase M, Cordes N (2007) Signalling via integrins: implications for cell survival and anticancer strategies. Biochim Biophys Acta 1775(1):163–180PubMedGoogle Scholar
  15. 15.
    Lemmon MA, Schlessinger J (2010) Cell signaling by receptor tyrosine kinases. Cell 141(7):1117–1134CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Gilmore AP (2005) Anoikis. Cell Death Differ 12(2):1473–1477CrossRefPubMedGoogle Scholar
  17. 17.
    Yamada KM, Even-Ram S (2002) Integrin regulation of growth factor receptors. Nat Cell Biol 4:E75–E76CrossRefPubMedGoogle Scholar
  18. 18.
    Borges E, Jan Y, Ruoslahti E (2000) Platelet-derived growth factor receptor βand vascular endothelial growth factor receptor2 bind to the β3 integrin through its extracellular domain. J Biol Chem 275:39867–39873CrossRefPubMedGoogle Scholar
  19. 19.
    Kruyt FA, Schuringa JJ (2010) Apoptosis and cancer stem cells: implications for apoptosis targeted therapy. Biochem Pharmacol 80:423–430CrossRefPubMedGoogle Scholar
  20. 20.
    An H, Kim JY, Oh E, Lee N, Cho Y, Seo JH (2015) Salinomycin promotes anoikis and decreases the CD44 +/CD24- stem-like population via inhibition of STAT3 activation in MDA-MB-231 cells. PLoS ONE 10(11):e0141919CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Harrison H, Farnie G, Howell SJ, Rock RE, Stylianou S, Brennan KR et al (2010) Regulation of breast cancer stem cell activity by signaling through the Notch4 receptor. Cancer Res 70(2):709–718CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Tang QL, Zhao ZQ, Li JC, Liang Y, Yin JQ, Zou CY et al (2011) Salinomycin inhibits osteosarcoma by targeting its tumor stem cells. Cancer Lett 311:113–121CrossRefPubMedGoogle Scholar
  23. 23.
    Fuji H, Honoki K, Tsujiuchi T, Kido A, Yoshitani K, Takakura Y (2009) Sphere-forming stem-like cell populations with drug resistance in human sarcoma cell lines. Int J Oncol 34(5):1381–1386Google Scholar
  24. 24.
    Lin Y, Chen G (2008) Embryoid body formation from human pluripotent stem cells in chemically defined E8 media. StemBook [Internet]. Cambridge (MA): Harvard Stem Cell Institute; 2008-Jun 1Google Scholar
  25. 25.
    Khwaja A, Rodriguez-Viciana P, Wennstrom S, Warne PH, Downward J (1997) Matrix adhesion and Ras transformation both activate a phosphoinositide 3-OH kinase and protein kinase B/Akt cellular survival pathway. EMBO J 16:2783–2793CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Schulze A, Lehmann K, Jefferies HB, McMahon M, Downward J (2001) Analysis of the transcriptional program induced by Raf in epithelial cells. Genes Dev 15:981–984CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Malanchi I, Santamaria-Martínez A, Susanto E, Peng H, Lehr HA, Delaloye JF et al (2011) Interactions between cancer stem cells and their niche govern metastatic colonization. Nature 481:85–89CrossRefPubMedGoogle Scholar
  28. 28.
    Langan RC, Mullinax JE, Raiji MT, Upham T, Summers T, Stojadinovic A et al (2013) Colorectal cancer biomarkers and the potential role of cancer stem cells. J Cancer 4(3):241–250CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Kerr JF, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–257CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Valentijn AJ, Zouq N, Gilmore AP (2004) Anoikis. Biochem Soc Trans 32(Pt3):421–425CrossRefPubMedGoogle Scholar
  31. 31.
    Reginato JM, Mills KR, Paulus JK, Lynch DK, Sgroi DC, Debnath J et al (2003) Integrins and EGFR coordinately regulate the pro-apoptotic protein Bim to prevent anoikis. Nat Cell Biol 5:733–740CrossRefPubMedGoogle Scholar
  32. 32.
    Galante JM, Mortenson MM, Bowles TL, Virudachalam S, Bold RJ (2009) ERK/BCL-2 pathway in the resistance of pancreatic cancer to anoikis. J Surg Res 152(1):18–25CrossRefPubMedGoogle Scholar
  33. 33.
    Miranti CK, Brugge JS (2002) Sensing the environment: a historical perspective on integrin signal transduction. Nat Cell Biol 4:E83–E90CrossRefPubMedGoogle Scholar
  34. 34.
    Morozevich GE, Kozlova NI, Susova OY, Karalkin PA, Berman AE (2015) Implication of α2β1 integrin in anoikis of MCF-7 human breast carcinoma cells. Biochemistry 80(1):97–103PubMedGoogle Scholar
  35. 35.
    Beauséjour M, Thibodeau S, Demers MJ, Bouchard V, Gauthier R, Beaulieu JF, Vachon PH (2013) Suppression of anoikis in human intestinal epithelial cells: differentiation state-selective roles of α2β1, α3β1, α5β1, and α6β4 integrins. BMC Cell Biol 14:53. CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Pouliot N, Nice EC, Burgess AW (2001) Laminin-10 mediates basal and EGF-stimulated motility of human colon carcinoma cells via alpha(3)beta(1) and alpha(6) beta(4) integrins. Exp Cell Res 266(1):1–10CrossRefPubMedGoogle Scholar
  37. 37.
    Ricono JM, Huang M, Barnes LA, Lau SK, Weis SM, Schlaepfer DD et al (2009) Specific cross-talk between epidermal growth factor receptor and Integrin αVβV promotes carcinoma cell invasion and metastasis. Cancer Res 69(4):1383–1391CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Alanko J, Mai A, Jacquemet G, Schauer K, Kaukonen R, Saari M et al (2015) Integrin endosomal signalling suppresses anoikis. Nat Cell Biol 17:1412–1421CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Yu X, Miyamoto S, Mekada E (2000) Integrin α2β1-dependent EGF receptor activation at cell-cell contact sites. J Cell Sci 113:2139–2147PubMedGoogle Scholar
  40. 40.
    Demers MJ, Thibodeau S, Noël D, Fujita N, Tsuruo T, Gauthier R, Arguin M, Vachon PH (2009) Intestinal epithelial cancer cell anoikis resistance: EGFR-mediated sustained activation of Src overrides Fak-dependent signaling to MEK/Erk and/or PI3-K/Akt-1. J Cell Biochem 107(4):639–654. CrossRefPubMedGoogle Scholar
  41. 41.
    Haenssen KK, Caldwell SA, Shahriari KS, Jackson SR, Whelan KA, Klein-Szanto AJ (2010) ErbB2 requires integrin alpha5 for anoikis resistance via Src regulation of receptor activity in human mammary epithelial cells. J Cell Sci 123(Pt 8):1373–1382CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Moro L, Dolce L, Cabodi S, Bergatto E, Boeri Erba E, Smeriglio M et al (2002) Integrin-induced epidermal growth factor (EGF) receptor activation requires c-Src and p130Cas and leads to phosphorylation of specific EGF receptor tyrosines. J Biol Chem 277(11):9405–9414CrossRefPubMedGoogle Scholar
  43. 43.
    Collins NL, Reginato MJ, Paulus JK, Sgroi DC, LaBaer J, Brugge JS (2005) G1/S cell cycle arrest provides anoikis resistance through erk-mediated bim suppression. Mol Cell Biol 25(12):5282–5291CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Cai Q, Yan L, Xu Y (2015) Anoikis resistance is a critical feature of highly aggressive ovarian cancer cells. Oncogene 34(25):3315–3324CrossRefPubMedGoogle Scholar
  45. 45.
    Cabodi S, Cabodi S, Moro L, Bergatto E, Boeri EE, Di Stefano P, Turco E et al (2004) Integrin regulation of epidermal growth factor (EGF) receptor and of EGF-dependent responses. Biochem Soc Trans 32:438–442CrossRefPubMedGoogle Scholar
  46. 46.
    Giancotti FG, Tarone G (2003) Positional control of cell fate through joint integrin/receptor protein kinase signaling. Annu Rev Cell Dev Biol 19:173–206CrossRefPubMedGoogle Scholar
  47. 47.
    Frisch SM, Schaller M, Cieply B (2013) Mechanisms that link the oncogenic epithelial-mesenchymal transition to suppression of anoikis. J Cell Sci 126:21–29CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Munoz P, Iliou MS, Esteller M (2012) Epigenetic alterations involved in cancer stem cell reprogramming. Mol Oncol 6:620–636CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Baccelli I, Trumpp A (2012) The evolving concept of cancer and metastasis stem cells. J Cell Biol 198:281–293CrossRefPubMedPubMedCentralGoogle Scholar
  50. 50.
    Grotenhuis BA, Wijnhoven BP, van Lanschot JJ (2012) Cancer stem cells and their potential implications for the treatment of solid tumors. J Surg Oncol 106:209–215CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Deblina Guha
    • 1
  • Taniya Saha
    • 1
  • Sayantan Bose
    • 1
  • Sreeparna Chakraborty
    • 1
  • Subhanki Dhar
    • 1
  • Poulami Khan
    • 1
  • Arghya Adhikary
    • 2
  • Tanya Das
    • 1
  • Gaurisankar Sa
    • 1
    Email author
  1. 1.Division of Molecular MedicineBose InstituteKolkataIndia
  2. 2.Center for Research in Nanoscience and Nanotechnology, Acharya Prafulla Chandra Roy Sikhsha PranganUniversity of CalcuttaSaltlake City, KolkataIndia

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