Skip to main content
  • Research Article
  • Published:

siRNA-mediated silencing of the 37/67-kDa high affinity laminin receptor in Hep3B cells induces apoptosis

Abstract

The laminin-binding protein, variously called the 37/67-kDa high affinity laminin receptor or p40, mediates the attachment of normal cells to the laminin network, and also has a role as a ribosomal protein. Over-expression of this protein has been strongly correlated with the metastatic phenotype. However, few studies have investigated the cellular consequence of the ablation of this gene’s expression. To address this issue, the expression of the 37/67-kDa high affinity laminin receptor was knocked out with several siRNA constructs via RNA interference in transformed liver (Hep3B) cells. In each case where the message was specifically ablated, apoptosis was induced, as determined by annexin V/propidium iodide staining, and by double staining with annexin V and an antibody directed against the 37/67-kDa high affinity laminin receptor. These results suggest that this protein plays a critical role in maintaining cell viability.

Abbreviations

GAPDH:

glyceraldehyde 3-phosphate dehydrogenase

LBP-p40:

laminin-binding protein precursor p40

LRP:

laminin receptor precursor

PrPc :

cellular prion protein

siRNA:

small interfering RNAs

37LBP:

37-kDa laminin binding protein

67LR:

67-kDa laminin receptor

References

  1. Lesot, H., Kuhl, U. and Mark, K.V. Isolation of a laminin-binding protein from muscle cell membranes. EMBO J. 2 (1983) 861–865.

    PubMed  CAS  Google Scholar 

  2. Malinoff, H.L. and Wicha, M.S. Isolation of a cell surface receptor protein for laminin from murine fibrosarcoma cells. J. Cell Biol. 96 (1983) 1475–1479.

    Article  PubMed  CAS  Google Scholar 

  3. Rao, N.C., Barsky, S.H., Terranova, V.P. and Liotta, L.A. Isolation of a tumor cell laminin receptor. Biochem. Biophys. Res. Commun. 111 (1983) 804–808.

    Article  PubMed  CAS  Google Scholar 

  4. Rao, C.N., Castronovo, V., Schmitt, M.C., Wewer, U.M., Claysmith, A.P., Liotta, L.A. and Sobel, M.E. Evidence for a precursor of the high-affinity metastasis-associated murine laminin receptor. Biochemistry 28 (1989) 7476–7486.

    Article  PubMed  CAS  Google Scholar 

  5. Yow, H.K., Wong, J.M., Chen, H.S., Lee, C.G., Davis, S., Steele, G.D., Jr. and Chen, L.B. Increased mRNA expression of a laminin-binding protein in human colon carcinoma: complete sequence of a full-length cDNA encoding the protein. Proc. Natl. Acad. Sci. USA 85 (1988) 6394–6398.

    Article  PubMed  CAS  Google Scholar 

  6. Tohgo, A., Takasawa, S., Munakata, H., Yonekura, H., Hayashi, N. and Okamoto, H. Structural determination and characterization of a 40-kDa protein isolated from rat 40 S ribosomal subunit. FEBS Lett. 340 (1994) 133–138.

    Article  PubMed  CAS  Google Scholar 

  7. Auth, D. and Brawerman, G. A 33-kDa polypeptide with homology to the laminin receptor: component of translation machinery. Proc. Natl. Acad. Sci. USA 89 (1992) 4368–4372.

    Article  PubMed  CAS  Google Scholar 

  8. Sato, M., Kinoshita, K., Kaneda, Y., Saeki, Y., Iwamatsu, A. and Tanaka, K. Analysis of nuclear localization of laminin binding protein precursor p40 (LBP/p40). Biochem. Biophys. Res. Commun. 229 (1996) 896–901.

    Article  PubMed  CAS  Google Scholar 

  9. Mecham, R.P., Hinek, A., Griffin, G.L., Senior, R.M. and Liotta, L.A. The elastin receptor shows structural and functional similarities to the 67-kDa tumor cell laminin receptor. J. Biol. Chem. 264 (1989) 16652–16657.

    PubMed  CAS  Google Scholar 

  10. McCaffery, P., Neve, R.L. and Drager, U.C. A dorso-ventral asymmetry in the embryonic retina defined by protein conformation. Proc. Natl. Acad. Sci. USA 87 (1990) 8570–8574.

    Article  PubMed  CAS  Google Scholar 

  11. Rieger, R., Edenhofer, F., Lasmezas, C.I. and Weiss, S. The human 37-kDa laminin receptor precursor interacts with the prion protein in eukaryotic cells. Nat. Med. 3 (1997) 1383–1388.

    Article  PubMed  CAS  Google Scholar 

  12. Gauczynski, S., Peyrin, J.M., Haik, S., Leucht, C., Hundt, C., Rieger, R., Krasemann, S., Deslys, J.P., Dormont, D., Lasmezas, C.I. and Weiss, S. The 37-kDa/67-kDa laminin receptor acts as the cell-surface receptor for the cellular prion protein. EMBO J. 20 (2001) 5863–5875.

    Article  PubMed  CAS  Google Scholar 

  13. Gauczynski, S., Nikles, D., El-Gogo, S., Papy-Garcia, D., Rey, C., Alban, S., Barritault, D., Lasmezas, C.I. and Weiss, S. The 37-kDa/67-kDa laminin receptor acts as a receptor for infectious prions and is inhibited by polysulfated glycanes. J. Infect. Dis. 194 (2006) 702–709.

    Article  PubMed  CAS  Google Scholar 

  14. Ludewigs, H., Zuber, C., Vana, K., Nikles, D., Zerr, I. and Weiss, S. Therapeutic approaches for prion disorders. Expert Rev. Anti. Infect. Ther. 5 (2007) 613–630.

    Article  PubMed  CAS  Google Scholar 

  15. Vana, K., Zuber, C., Nikles, D. and Weiss, S. Novel aspects of prions, their receptor molecules, and innovative approaches for TSE therapy. Cell. Mol. Neurobiol. 27 (2007)107–128.

    Article  PubMed  CAS  Google Scholar 

  16. Zuber, C., Ludewigs, H. and Weiss, S. Therapeutic approaches targeting the prion receptor LRP/LR. Vet. Microbiol. 123 (2007) 387–393.

    Article  PubMed  CAS  Google Scholar 

  17. Wang, K.S., Kuhn, R.J., Strauss, E.G., Ou, S. and Strauss, J.H. High-affinity laminin receptor is a receptor for Sindbis virus in mammalian cells. J. Virol. 66 (1992) 4992–5001.

    PubMed  CAS  Google Scholar 

  18. Thepparit, C. and Smith, D.R. Serotype-specific entry of dengue virus into liver cells: identification of the 37-kilodalton/67-kilodalton high-affinity laminin receptor as a dengue virus serotype 1 receptor. J. Virol. 78 (2004) 12647–12656.

    Article  PubMed  CAS  Google Scholar 

  19. Akache, B., Grimm, D., Pandey, K., Yant, S.R., Xu, H. and Kay, M.A. The 37/67-kilodalton laminin receptor is a receptor for adeno-associated virus serotypes 8, 2, 3, and 9. J. Virol. 80 (2006) 9831–9836.

    Article  PubMed  CAS  Google Scholar 

  20. Coggin, J.H. Jr., Barsoum, A.L. and Rohrer, J.W. 37 kiloDalton oncofetal antigen protein and immature laminin receptor protein are identical, universal T-cell inducing immunogens on primary rodent and human cancers. Anticancer Res. 19 (1999) 5535–5542.

    PubMed  CAS  Google Scholar 

  21. Zelle-Rieser, C., Barsoum, A.L., Sallusto, F., Ramoner, R., Rohrer, J.W., Holtl, L., Bartsch, G., Coggin, J.J. and Thurnher, M. Expression and immunogenicity of oncofetal antigen-immature laminin receptor in human renal cell carcinoma. J. Urol. 165 (2001) 1705–1709.

    Article  PubMed  CAS  Google Scholar 

  22. Castronovo, V. Laminin receptors and laminin-binding proteins during tumor invasion and metastasis. Invasion Metastasis 13 (1993) 1–30.

    PubMed  CAS  Google Scholar 

  23. Castronovo, V., Claysmith, A.P., Barker, K.T., Cioce, V., Krutzsch, H.C. and Sobel, M.E. (1991). Biosynthesis of the 67 kDa high affinity laminin receptor. Biochem. Biophys. Res. Commun. 177 (1991) 177–183.

    Article  PubMed  CAS  Google Scholar 

  24. Buto, S., Tagliabue, E., Ardini, E., Magnifico, A., Ghirelli, C., van den Brule, F., Castronovo, V., Colnaghi, M.I., Sobel, M.E. and Menard, S. Formation of the 67-kDa laminin receptor by acylation of the precursor. J. Cell. Biochem. 69 (1998) 244–251.

    Article  PubMed  CAS  Google Scholar 

  25. Narumi, K., Inoue, A., Tanaka, M., Isemura, M., Shimo-Oka, T., Abe, T., Nukiwa, T. and Satoh, K. Inhibition of experimental metastasis of human fibrosarcoma cells by anti-recombinant 37-kDa laminin binding protein antibody. Jpn. J. Cancer Res. 90 (1999) 425–431.

    PubMed  CAS  Google Scholar 

  26. Kaneda, Y., Kinoshita, K., Sato, M., Saeki, Y., Yamada, R., Wataya-Kaneda, M., and Tanaka, K. The induction of apoptosis in HeLa cells by the loss of LBP-p40. Cell Death Differ. 5 (1998) 20–28.

    Article  PubMed  CAS  Google Scholar 

  27. Leucht, C., Simoneau, S., Rey, C., Vana, K., Rieger, R., Lasmezas, C.I. and Weiss, S. The 37 kDa/67 kDa laminin receptor is required for PrP(Sc) propagation in scrapie-infected neuronal cells. EMBO Rep. 4 (2003) 290–295.

    Article  PubMed  CAS  Google Scholar 

  28. Leucht, C., Vana, K., Renner-Muller, I., Dormont, D., Lasmezas, C.I., Wolf, E. and Weiss, S. Knock-down of the 37-kDa/67-kDa laminin receptor in mouse brain by transgenic expression of specific antisense LRP RNA. Transgenic Res. 13 (2004) 81–85.

    Article  PubMed  CAS  Google Scholar 

  29. Ardini, E., Pesole, G., Tagliabue, E., Magnifico, A., Castronovo, V., Sobel, M.E., Colnaghi, M.I. and Menard, S. The 67-kDa laminin receptor originated from a ribosomal protein that acquired a dual function during evolution. Mol. Biol. Evol. 15 (1998) 1017–1025.

    PubMed  CAS  Google Scholar 

  30. Knowles B.B., Howe, C.C. and Aden D.P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science 209 (1980) 497–499.

    Article  PubMed  CAS  Google Scholar 

  31. Elbashir, S.M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K. and Tuschl, T. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411 (2001) 494–498.

    Article  PubMed  CAS  Google Scholar 

  32. Hannon, G.J. RNA interference. Nature 418 (2002) 244–251.

    Article  PubMed  CAS  Google Scholar 

  33. Hurtado, A., Tseng, J.C. and Meruelo, D. Gene therapy that safely targets and kills tumor cells throughout the body. Rejuvenation Res. 9 (2006) 36–44.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Duncan R. Smith.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Susantad, T., Smith, D.R. siRNA-mediated silencing of the 37/67-kDa high affinity laminin receptor in Hep3B cells induces apoptosis. Cell Mol Biol Lett 13, 452–464 (2008). https://doi.org/10.2478/s11658-008-0017-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-008-0017-6

Key words