Biochemistry (Moscow)

, Volume 79, Issue 6, pp 524–530 | Cite as

Study of Wnt2 secreted by A-549 cells in paracrine activation of β-catenin in co-cultured mesenchymal stem cells

  • N. S. Petrov
  • B. V. PopovEmail author


The canonical Wnt signal pathway is a key regulator of self-renewal and cell fate determination in various types of stem cells. The total pool of β-catenin consists of two different forms: the signaling form of the protein transmits the Wnt signals from the cell membrane to the target genes, whereas the membrane β-catenin is involved in formation of cell-to-cell contact at cadherin junctions. Earlier we developed an in vitro model of epithelial differentiation of mesenchymal stem cells (MSCs) co-cultured with epithelial A-549 cells. The purpose of the present work was to study the role of Wnt2 secreted by the A-549 cells in paracrine induction of β-catenin in co-cultured MSCs. Using the somatic gene knockdown technique, we obtained A-549 cell cultures with down-regulated WNT2. The MSCs co-cultured with the control A-549 cells displayed an increase in the levels of total cellular and signaling β-catenin and transactivation of a reporter construction containing the Lef/Tcf protein family binding sites. In contrast, β-catenin was not induced in the MSCs co-cultured with the A-549 cells with down-regulated WNT2 expression, but the total protein level was increased. We suggest that Wnt2 secreted by A-549 cells induces in co-cultured MSCs the Wnt/β-catenin signaling pathway, whereas the associated increase in total β-catenin level should be due to another mechanism.

Key words

mesenchymal stem cells Wnt/β-catenin signaling pathway 



active (signaling) β-catenin


mesenchymal stem cells


total β-catenin


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Schuijers, J., and Clevers, H. (2012) EMBO J., 31, 2685–2696.PubMedCentralPubMedCrossRefGoogle Scholar
  2. 2.
    Nusse, R., and Varmus, H. (2012) EMBO J., 31, 2670–2684.PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Behrens, J., von Kries, J. P., Kuhl, M., Bruhn, L., Wedlich, D., Grosschedl, R., and Birchmeier, W. (1996) Nature, 382, 638–642.PubMedCrossRefGoogle Scholar
  4. 4.
    Huber, O., Korn, R., McLaughlin, J., Ohsugi, M., Herrmann, B. G., and Kemler, R. (1996) Mech. Dev., 59, 3–10.PubMedCrossRefGoogle Scholar
  5. 5.
    Lyashenko, N., Winter, M., Migliorini, D., Biechele, T., Moon, R. T., and Hartmann, C. (2011) Nat. Cell Biol., 13, 753–761.PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Reya, T., and Clevers, H. (2005) Nature, 434, 843–850.PubMedCrossRefGoogle Scholar
  7. 7.
    Ying, Q. L., Wray, J., Nichols, J., Batlle-Morera, L., Doble, B., Woodgett, J., Cohen, P., and Smith, A. (2008) Nature, 453, 519–523.PubMedCrossRefGoogle Scholar
  8. 8.
    Van Es, J. H., van Gijn, M. E., Riccio, O., van den Born, M., Vooijs, M., Begthel, H., Cozijnsen, M., Robine, S., Winton, D. J., Radtke, F., and Clevers, H. (2005) Nature, 435, 959–963.PubMedCrossRefGoogle Scholar
  9. 9.
    Alonso, L., and Fuchs, E. (2003) Proc. Natl. Acad. Sci. USA, 100,Suppl. 1, 11830–11835.PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Ling, L., Nurcombe, V., and Cool, S. M. (2009) Gene, 433, 1–7.PubMedCrossRefGoogle Scholar
  11. 11.
    Gottardi, C. J., and Gumbiner, B. M. (2004) J. Cell Biol., 167, 339–349.PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Heuberger, J., and Birchmeier, W. (2010) Cold Spring Harb. Perspect. Biol., 2, a002915.PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Kimelman, D., and Xu, W. (2006) Oncogene, 25, 7482–7491.PubMedCrossRefGoogle Scholar
  14. 14.
    Huber, A. H., and Weis, W. I. (2001) Cell, 105, 391–402.PubMedCrossRefGoogle Scholar
  15. 15.
    Korswagen, H. C., Herman, M. A., and Clevers, H. C. (2000) Nature, 406, 527–532.PubMedCrossRefGoogle Scholar
  16. 16.
    Liu, J., Phillips, B. T., Amaya, M. F., Kimble, J., and Xu, W. (2008) Dev. Cell, 14, 751–761.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Liu, C., Li, Y., Semenov, M., Han, C., Baeg, G. H., Tan, Y., Zhang, Z., Lin, X., and He, X. (2002) Cell, 108, 837–847.PubMedCrossRefGoogle Scholar
  18. 18.
    Papkoff, J., Rubinfeld, B., Schryver, B., and Polakis, P. (1996) Mol. Cell Biol., 16, 2128–21234.PubMedCentralPubMedGoogle Scholar
  19. 19.
    Hart, M. J., de los Santos, R., Albert, I. N., Rubinfeld, B., and Polakis, P. (1998) Curr. Biol., 8, 573–581.PubMedCrossRefGoogle Scholar
  20. 20.
    Wu, D., and Pan, W. (2010) Trends Biochem. Sci., 35, 161–168.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Kishida, S., Yamamoto, H., Ikeda, S., Kishida, M., Sakamoto, I., Koyama, S., and Kikuchi, A. (1998) J. Biol. Chem., 27, 10823–10826.CrossRefGoogle Scholar
  22. 22.
    Sakanaka, C., Weiss, J. B., and Williams, L. T. (1998) Proc. Natl. Acad. Sci. USA, 95, 3020–3023.PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Su, Y., Fu, C., Ishikawa, S., Stella, A., Kojima, M., Shitoh, K., Schreiber, E. M., Day, B. W., and Liu, B. (2008) Mol. Cell, 32, 652–661.PubMedCrossRefGoogle Scholar
  24. 24.
    Korinek, V., Barker, N., Morin, P. J., van Wichen, D., de Weger, R., Kinzler, K. W., Vogelstein, B., and Clevers, H. (1997) Science, 275, 1784–1787.PubMedCrossRefGoogle Scholar
  25. 25.
    Morin, P. J., Sparks, A. B., Korinek, V., Barker, N., Clevers, H., Vogelstein, B., and Kinzler, K. W. (1997) Science, 275, 1787–1790.PubMedCrossRefGoogle Scholar
  26. 26.
    Li, V. S., Ng, S. S., Boersema, P. J., Low, T. Y., Karthaus, W. R., Gerlach, J. P., Mohammed, S., Heck, A. J., Maurice, M. M., Mahmoudi, T., and Clevers, H. (2012) Cell, 149, 1245–1256.PubMedCrossRefGoogle Scholar
  27. 27.
    Hernandez, A. R., Klein, A. M., and Kirschner, M. W. (2012) Science, 338, 1337–1340.PubMedCrossRefGoogle Scholar
  28. 28.
    Nguyen, H., Rendl, M., and Fuchs, E. (2006) Cell, 127, 171–183.PubMedCrossRefGoogle Scholar
  29. 29.
    Archbold, H. C., Yang, Y. X., Chen, L., and Cadigan, K. M. (2012) Acta Physiol. (Oxford), 204, 74–109.CrossRefGoogle Scholar
  30. 30.
    Popov, B. V., Serikov, V. B., Petrov, N. S., Izusova, T. V., Gupta, N., and Matthay, M. A. (2007) Tissue Eng., 13, 2441–2450.PubMedCrossRefGoogle Scholar
  31. 31.
    You, L., He, B., Xu, Z., Uematsu, K., Mazieres, J., Fujii, N., Mikami, I., Reguart, N., McIntosh, J. K., Kashani-Sabet, M., McCormick, F., and Jablons, D. M. (2004) Cancer Res., 64, 5385–5389.PubMedCrossRefGoogle Scholar
  32. 32.
    Stambolic, V., Ruel, L., and Woodgett, J. R. (1996) Curr. Biol., 6, 1664–1668.PubMedCrossRefGoogle Scholar
  33. 33.
    Ray, K. P., Farrow, S., Daly, M., Talabot, F., and Searle, N. (1997) Biochem. J., 328, 707–715.PubMedCentralPubMedGoogle Scholar
  34. 34.
    Brummelkamp, T. R., Bernards, R., and Agami, R. (2002) Science, 296, 550–553.PubMedCrossRefGoogle Scholar
  35. 35.
    Aberle, H., Bauer, A., Stappert, J., Kispert, A., and Kemler, R. (1997) EMBO J., 16, 3797–3804.PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Popov, B. V., Watt, S. W., Rozanov, Iu. M., and Chang, L. S. (2010) Mol. Biol., 44, 323–334.Google Scholar
  37. 37.
    Staal, F. J., Noort, M. M., Strous, G. J., and Clevers, H. C. (2002) EMBO Rep., 3, 63–68.PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Fuxe, J., Vincent, T., and Garcia de Herreros, A. (2010) Cell Cycle, 9, 2363–2374.PubMedCrossRefGoogle Scholar
  39. 39.
    Krejci, P., Aklian, A., Kaucka, M., Sevcikova, E., Prochazkova, J., Masek, J. K., Mikolka, P., Pospisilova, T., Spoustova, T., Weis, M., Paznekas, W. A., Wolf, J. H., Gutkind, J. S., Wilcox, W. R., Kozubik, A., Jabs, E. W., Bryja, V., Salazar, L., Vesela, I., and Balek, L. (2012) PLoS One, 7, e35826.PubMedCentralPubMedCrossRefGoogle Scholar
  40. 40.
    Palsgaard, J., Emanuelli, B., Winnay, J. N., Sumara, G., Karsenty, G., and Kahn, C. R. (2012) J. Biol. Chem., 287, 12016–12026.PubMedCentralPubMedCrossRefGoogle Scholar
  41. 41.
    Knudsen, E. S., and Wang, J. Y. J. (1996) J. Biol. Chem., 271, 8313–8320.PubMedCrossRefGoogle Scholar
  42. 42.
    Roman-Gomez, J., Cordeu, L., Agirre, X., Jimenez-Velasco, A., San Jose-Eneriz, E., Garate, L., Calasanz, M. J., Heiniger, A., Torres, A., and Prosper, F. (2007) Blood, 109, 3462–3469.PubMedCrossRefGoogle Scholar
  43. 43.
    Lam, A. P., Flozak, A. S., Russell, S., Wei, J., Jain, M., Mutlu, G. M., Budinger, G. R., Feghali-Bostwick, C. A., Varga, J., and Gottardi, C. J. (2011) Am. J. Respir. Cell Mol. Biol., 45, 915–922.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2014

Authors and Affiliations

  1. 1.Institute of CytologyRussian Academy of SciencesSt. PetersburgRussia

Personalised recommendations