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Implication of Integrin α2β1 in Proliferation and Invasion of Human Breast Carcinoma and Melanoma Cells: Noncanonical Function of Akt Protein Kinase

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Abstract

Blocking the expression of integrin α2β1, which was accomplished by transduction of α2-specific shRNA, resulted in significant inhibition of proliferation and clonal activity in human MCF-7 breast carcinoma and SK-Mel-147 melanoma cells. Along with these changes, deprivation of α2β1 caused a sharp decrease in melanoma cell invasion in vitro. Analysis of integrin-mediating signal pathways that control cell behavior revealed a significant increase in activity of Akt protein kinase in response to depletion of α2β1. The increase in Akt activity that accompanies a suppressive effect on cell invasion contradicts well-known Akt function aimed at stimulation of tumor progression. This contradiction could be explained by the “reversed” (noncanonical) role played by Akt in some cells that consists in suppression rather than promotion of invasive phenotype. To test this suggestion, the effects of Akt inhibitors on invasive activity of SK-Mel-147 cells were investigated. If the above suggestion is true, then inhibition of Akt in cells depleted of α2β1 should result in the restoration of their invasive activity. It appeared that treatment with LY294002, which inhibits all Akt isoforms (Akt1, Akt2, Akt3), not only failed to restore the invasive phenotype of melanoma cells but further attenuated their invasive activity. However, treatment of the cells with an Akt1-specific inhibitor significantly increased their invasion. Thus, the stimulating effect of α2β1 integrin on invasion of melanoma cells is realized through a mechanism based on inhibition of one of the Akt isoforms, which in these cells exhibits a noncanonical function consisting in suppression of invasion.

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Abbreviations

ECM:

extracellular matrix

EdU:

5-ethynyl-2′-deoxyuridine

References

  1. Guo, W., and Giancotti, F. G. (2004) Integrin signalling during tumour progression, Nat. Rev. Mol. Cell. Biol., 10, 816–826.

    Article  CAS  Google Scholar 

  2. Hehlgans, S., Haase, M., and Cordes, N. (2007) Signalling via integrins: implications for cell survival and anticancer strategies, Biochim. Biophys. Acta, 1775, 163–180.

    PubMed  CAS  Google Scholar 

  3. Desgrosellier, J. S., and Cheresh, D. A. (2010) Integrins in cancer: biological implications and therapeutic opportunities, Nat. Rev. Cancer, 10, 9–22.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Kuphal, S., Bauer, R., and Bosserhoff, A. K. (2005) Integrin signaling in malignant melanoma, Cancer Metastasis Rev., 24, 195–222.

    Article  PubMed  CAS  Google Scholar 

  5. Haenssen, K. K., Caldwell, S. A., Shahriari, K. S., Jackson, S. R., Whelan, K. A., Klein-Szanto, A. J., and Reginato, M. J. (2010) ErbB2 requires integrin alpha 5 for anoikis resistance via Src regulation of receptor activity in human mammary epithelial cells, J. Cell Sci., 123, 1373–1382.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Bai, J., Zhang, J., Wu, J., Shen, L., Zeng, J., Ding, J., Wu, Y., Gong, Z., Li, A., Xu, S., Zhou, J., and Li, G. (2010) JWA regulates melanoma metastasis by integrin alphaVbeta 3 signaling, Oncogene, 29, 1227–1237.

    Article  PubMed  CAS  Google Scholar 

  7. Nam, E. H., Lee, Y., Moon, B., Lee, J. W., and Kim, S. (2015) Twist1 and AP-1 cooperatively upregulate integrin α5 expression to induce invasion and the epithelial–mes-enchymal transition, Carcinogenesis, 36, 327–337.

    Article  PubMed  CAS  Google Scholar 

  8. Tran, T., Barlow, B., O’Rear, L., Jarvis, B., Li, Z., Dickeson, K., Dupont, W., and Zutter, M. (2011) Loss of the α2β1 integrin alters human papilloma virus-induced squamous carcinoma progression in vivo and in vitro, PLoS One, 6, e26858.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Girotti, M. R., Fernandez, M., Lopez, J. A., Camafeita, E., Fernandez, E. A., Albar, J. P., Benedetti, L. G., Valacco, M. P., Brekken, R. A., Podhajcer, O. L., and Llera, A. S. (2011) SPARC promotes cathepsin B-mediat-ed melanoma invasiveness through a collagen I/α2β1 inte-grin axis, J. Invest. Dermatol., 131, 2438–2447.

    Article  PubMed  CAS  Google Scholar 

  10. Burnier, J. V., Wang, N., Michel, R. P., Hassanain, M., Li, S., Lu, Y., Metrakos, P., Antecka, E., Burnier, M. N., Ponton, A., Gallinger, S., and Brodt, P. (2011) Type IV col-lagen-initiated signals provide survival and growth cues required for liver metastasis, Oncogene, 30, 3766–3783.

    Article  PubMed  CAS  Google Scholar 

  11. Li, X., Ishihara, S., Yasuda, M., Nishioka, T., Mizutani, T., Ishikawa, M., Kawabata, K., Shirato, H., and Haga, H. (2013) Lung cancer cells that survive ionizing radiation show increased integrin α2β1-and EGFR-dependent inva-siveness, PLoS One, 8, e70905.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  12. Ramirez, N. E., Zhang, Z., Madamanchi, A., Boyd, K. L., O’Rear, L. D., Nashabi, A., Li, Z., Dupont, W. D., Zijlstra, A., and Zutter, M. M. (2011) The α2β1 integrin is a metas-tasis suppressor in mouse models and human cancer, J. Clin. Invest., 121, 226–237.

    Article  PubMed  CAS  Google Scholar 

  13. Ferraro, A., Mourtzoukou, D., Kosmidou, V., Avlonitis, S., Kontogeorgos, G., Zografos, G., and Pintzas, A. (2013) EZH2 is regulated by ERK/AKT and targets integrin alpha2 gene to control epithelial–mesenchymal transition and anoikis in colon cancer cells, Int. J. Biochem. Cell Biol., 45, 243–254.

    Article  PubMed  CAS  Google Scholar 

  14. Naci, D., Vuori, K., and Aoudjit, F. (2015) Alpha2beta1 integrin in cancer development and chemoresistance, Semin. Cancer Biol., 35, 145–153.

    Article  PubMed  CAS  Google Scholar 

  15. Guo, Y. S., Zhao, R., Ma, J., Cui, W., Sun, Z., Gao, B., He, S., Han, Y. H., Fan, J., Yang, L., Tang, J., and Luo, Z. J. (2014) βig-h3 promotes human osteosarcoma cells metastasis by interacting with integrin α2β1 and activating PI3K signaling pathway, PLoS One, 9, e90220.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  16. Naci, D., Azreq, M. A., Chetoui, N., Lauden, L., Sigaux, F., Charron, D., Al-Daccak, R., and Aoudjit, F. (2012) α2β1 integrin promotes chemoresistance against doxoru-bicin in cancer cells through extracellular signal-regulated kinase (ERK), J. Biol. Chem., 287, 17065–17076.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Mannava, S., Omilian, A. R., Wawrzyniak, J. A., Fink, E. E., Zhuang, D., Miecznikowski, J. C., Marshall, J. R., Soengas, M. S., Sears, R. C., Morrison, C. D., and Nikiforov, M. A. (2012) PP2A-B56a controls oncogene-induced senescence in normal and tumor human melanocytic cells, Oncogene, 31, 1484–1492.

    Article  PubMed  CAS  Google Scholar 

  18. Morozevich, G. E., Kozlova, N. I., Ushakova, N. A., Preobrazhenskaya, M. E., and Berman, A. E. (2012) Integrin α5β1 simultaneously controls EGFR-dependent proliferation and Akt-dependent pro-survival signaling in epidermoid carcinoma cells, Aging (Albany NY), 4, 368–374.

    Article  CAS  Google Scholar 

  19. Morozevich, G. E., Kozlova, N. I., Cheglakov, I. B., Ushakova, N. A., and Berman, A. E. (2009) Integrin α5β1 controls invasion of human breast carcinoma cells by direct and indirect modulation of MMP-2 collagenase activity, Cell Cycle, 14, 2219–2225.

    Article  Google Scholar 

  20. Morozevich, G. E., Kozlova, N. I., Susova, O. Y., Karalkin, P. A., and Berman, A. E. (2015) Implication of α2β1 integrin in anoikis of MCF-7 breast carcinoma cells, Biochemistry (Moscow), 80, 97–103.

    Article  CAS  Google Scholar 

  21. Krueger, J. S., Keshamouni, V. G., Atanaskova, N., and Reddy, K. B. (2001) Temporal and quantitative regulation of mitogen-activated protein kinase (MAPK) modulates cell motility and invasion, Oncogene, 20, 4209–4218.

    Article  PubMed  CAS  Google Scholar 

  22. Okayama, H. (2012) Cdc6: a trifunctional AAA+ ATPase that plays a central role in controlling the G(1)-S transition and cell survival, J. Biochem., 152, 297–303.

    Article  PubMed  CAS  Google Scholar 

  23. Benetatos, L., Vartholomatos, G., and Hatzimichael, E. (2014) Polycomb group proteins and MYC: the cancer con-nection, Cell. Mol. Life Sci., 71, 257–269.

    Article  PubMed  CAS  Google Scholar 

  24. Toulany, M., and Rodemann, H. P. (2016) Phosphatidylinositol 3-kinase/Akt signaling as a key medi-ator of tumor cell responsiveness to radiation, Semin. Cancer Biol., 35, 180–190.

    Article  CAS  Google Scholar 

  25. Safdari, Y., Khalili, M., Ebrahimzadeh, M. A., Yazdani, Y., and Farajnia, S. (2015) Natural inhibitors of PI3K/AKT signaling in breast cancer: emphasis on newly-discovered molecular mechanisms of action, Pharmacol. Res., 93, 1–10.

    Article  PubMed  CAS  Google Scholar 

  26. Toker, A., and Yoeli-Lerner, M. (2006) Akt signaling and cancer: surviving but not moving on, Cancer Res., 66, 3963–3966.

    Article  PubMed  CAS  Google Scholar 

  27. Irie, H. Y., Pearline, R. V., Grueneberg, D., Hsia, M., Ravichandran, P., Kothari, N., Natesan, S., and Brugge, J. S. (2005) Distinct roles of Akt1 and Akt2 in regulating cell migration and epithelial–mesenchymal transition, J. Cell Biol., 19, 1023–1034.

    Article  CAS  Google Scholar 

  28. Dyce, O. H., Ziober, A. F., Weber, R. S., Miyazaki, K., Khariwala, S. S., Feldman, M., and Ziober, B. L. (2002) Integrins in head and neck squamous cell carcinoma inva-sion, Laryngoscope, 112, 2025–2032.

    Article  PubMed  CAS  Google Scholar 

  29. Sottnik, J. L., Daignault-Newton, S., Zhang, X., Morrissey, C., Hussain, M. H., Keller, E. T., and Hall, C. L. (2013) Integrin alpha2beta1 (α2β1) promotes prostate cancer skeletal metastasis, Clin. Exp. Metastasis, 30, 569–578.

    Article  PubMed  CAS  Google Scholar 

  30. Yoshimura, K., Meckel, K. F., Laird, L. S., Chia, C. Y., Park, J. J., Olino, K. L., Tsunedomi, R., Harada, T., Iizuka, N., Hazama, S., Kato, Y., Keller, J. W., Thompson, J. M., Chang, F., Romer, L. H., Jain, A., Iacobuzio-Donahue, C., Oka, M., Pardoll, D. M., and Schulick, R. D. (2009) Integrin alpha2 mediates selective metastasis to the liver, Cancer Res., 69, 7320–7328.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Choi, J. A., Jung, Y. S., Kim, J. Y., Kim, H. M., and Lim, I. K. (2016) Inhibition of breast cancer invasion by TIS21/BTG2/Pc3-Akt1-Sp1-Nox4 pathway targeting actin nucleators, mDia genes, Oncogene, 35, 83–93.

    Article  PubMed  CAS  Google Scholar 

  32. Hutchinson, J. N., Jin, J., Cardiff, R. D., Woodgett, J. R., and Muller, W. J. (2004) Activation of Akt-1 (PKB-alpha) can accelerate ErbB-2-mediated mammary tumorigenesis but suppresses tumor invasion, Cancer Res., 64, 3171–3178.

    Article  PubMed  CAS  Google Scholar 

  33. Arboleda, M. J., Lyons, J. F., Kabbinavar, F. F., Bray, M. R., Snow, B. E., Ayala, R., Danino, M., Karlan, B. Y., and Slamon, D. J. (2003) Overexpression of Akt2/protein kinase Bbeta leads to up-regulation of beta1 integrins, increased invasion, and metastasis of human breast and ovarian cancer cells, Cancer Res., 63, 196–206.

    PubMed  CAS  Google Scholar 

  34. Riggio, M., Perrone, M. C., Polo, M. L., Rodriguez, M. J., May, M., Abba, M., Lanari, C., and Novaro, V. (2017) AKT1 and AKT2 isoforms play distinct roles during breast cancer progression through the regulation of specific down-stream proteins, Sci. Rep., 7; doi: 10.1038/srep44244.

  35. Rao, G., Pierobon, M., Kim, I. K., Hsu, W. H., Deng, J., Moon, Y.-W., Petricoin, E. F., Zhang, Y. W., Wang, Y., and Giaccone, G. (2017) Inhibition of AKT1 signaling pro-motes invasion and metastasis of non-small cell lung cancer cells with K-ras or EGFR mutations, Sci. Rep., 7; doi: 10.1038/s41598-017-06128-9.

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Authors and Affiliations

  1. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Sciences, 119121, Moscow, Russia

    N. I. Kozlova, G. E. Morozevich, N. A. Ushakova & A. E. Berman

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  1. N. I. Kozlova
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  2. G. E. Morozevich
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  3. N. A. Ushakova
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  4. A. E. Berman
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Correspondence to A. E. Berman.

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Published in Russian in Biokhimiya, 2018, Vol. 83, No. 6, pp. 939-948.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM18-007, April 9, 2018.

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Kozlova, N.I., Morozevich, G.E., Ushakova, N.A. et al. Implication of Integrin α2β1 in Proliferation and Invasion of Human Breast Carcinoma and Melanoma Cells: Noncanonical Function of Akt Protein Kinase. Biochemistry Moscow 83, 738–745 (2018). https://doi.org/10.1134/S0006297918060111

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  • DOI: https://doi.org/10.1134/S0006297918060111

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