Skip to main content
SpringerLink
Log in

Quantitative immunofluorescence estimation of Pgp expression in human solid tumors by flow cytometry

  • Pharmaceutical Chemistry
  • Published:
Moscow University Chemistry Bulletin Aims and scope

Abstract

An immunofluorescence assay that enables the quantitative detection of the level and intensity of the expression of multidrug resistance marker Pgp in human solid tumors using flow cytometry and monoclonal antibody (clone 17F9, BD Pharmingen) was developed and adapted for clinical assessment. The analysis includes the following relevant task-specific stages. Single-cell suspension can be prepared from both surgical biopsy native tumor tissues and tumor specimens fixed in 4% formaldehyde. The assay allows one to verify results, even after the tumor material has been stored for 2 years. The cost of analysis can be decreased by increasing the period of cell incubation with antibodies and reducing the antibody concentration. The possibility of analyzing minimal tumor samples, e.g., endoscopic biopsy specimens, by decreasing the concentration of cells incubated with antibodies to 100000 cells/mL and the number of cells used in the flow cytometer to 1000 has also been shown. Despite its continuance, the developed assay does not require any changes in the investigator’s work schedule.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Widmer, N., Colombo, S., Buclin, T., and Decosterd, L.A., Blood, 2003, vol. 102, no. 3, p. 1142.

    Article  CAS  Google Scholar 

  2. Radujkovic, A., Schad, M., Topaly, J., Veldwijk, M.R., Laufs, S., Schultheis, B.S., Jauch, A., Melo, J.V., Fruehauf, S., and Zeller, W.J., Leukemia, 2005, vol. 19, no. 7, p. 1198.

    Article  CAS  Google Scholar 

  3. Nakanishi, T., Shiozawa, K., Hassel, B.A., and Ross, D.D., Blood, 2006, vol. 108, no. 2. p. 678.

    Article  CAS  Google Scholar 

  4. Yang, C.H., Huang, C.J., Yang, C.S., Chu, Y.C., Cheng, A.L., Whang-Peng, J., and Yang, P.C., Cancer Res., 2005, vol. 65, no. 15, p. 6943.

    Article  CAS  Google Scholar 

  5. Nagashima, S., Soda, H., Oka, M., Kitazaki, T., Shiozawa, K., Nakamura, Y., Takemura, M., Yabuuchi, H., Fukuda, M., Tsukamoto, K., and Kohno, S., Cancer Chemother. Pharmacol., 2006, vol. 58, no. 5, p. 594.

    Article  CAS  Google Scholar 

  6. Leschziner, G.D., Andrew, T., Pirmohamed, M., and Johnson, M.R., Pharmacogenomics J., 2007, vol. 7, no. 3, p. 154.

    Article  CAS  Google Scholar 

  7. Higgins, C.F., Nature., 2007, vol. 446, no 7137, p. 749.

    Article  CAS  Google Scholar 

  8. Zhao, Y., Yu, L., Lou, F., Wang, Q., and Pu, J., Zhonghua Nei Ke Za Zhi, 1999, vol. 38 no. 11, p. 760.

    CAS  Google Scholar 

  9. Schvarzanbach, H., Anal. Biochim., 2002, vol. 308, no. 1, p. 26.

    Article  Google Scholar 

  10. Parker, R.L., Huntsman, D.G., Lesack, D.W., Cupples, J.B., Grant, D.R., Akbari, M., and Gilks, C.B., Am. J. Clin. Pathol., 2002, vol. 117, no 5, p. 723.

    Article  Google Scholar 

  11. Leonessa, F., and Clarke, R., Endocr. Relat. Cancer., 2003, vol. 10, no. 1,. p. 43.

    Article  CAS  Google Scholar 

  12. Roy, S., Kenny, E., Kennedy, S., Larkin, A., Ballot, J., Perez, De Villareal M., Vrown, J., and O’Driscoll L., Anticancer Res., 2007, vol. 27, no. 3A, p. 1325.

    CAS  Google Scholar 

  13. Kreisholt, J., Sorensen, M., Jensen, P.B., Nielsen, B.S., Andersen, C.B., and Sehested, M., Br., J. Cancer., 1998, vol. 77, no. 9, p. 1469.

    Article  CAS  Google Scholar 

  14. Peng, Z.M., Luo, J., Wang, W.B., Wang, X.H., Chen, J.H., and Lan S.M., Ai Zheng., 2004, vol. 23, no. 8, p. 963.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Federal State Budgetary Institution Blokhin “Russian Cancer Research Center”, under the Russian Academy of Medical Sciences, Moscow, Russia

    T. A. Bogush, M. V. Tikhomirov, E. A. Dudko, M. N. Sinitsyna, R. J. Ramanauskaite, B. E. Polotsky, S. A. Tjulandin & M. I. Davydov

Authors
  1. T. A. Bogush
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. V. Tikhomirov
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. E. A. Dudko
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. N. Sinitsyna
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R. J. Ramanauskaite
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. B. E. Polotsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S. A. Tjulandin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. M. I. Davydov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. A. Bogush.

Additional information

Original Russian Text © T.A. Bogush, M.V. Tikhomirov, E.A. Dudko, M.N. Sinitsyna, R.J. Ramanauskaite, B.E. Polotsky, S.A. Tjulandin, M.I. Davydov, 2012, published in Vestnik Moskovskogo Universiteta. Khimiya, 2012, No. 3, pp. 207–215.

About this article

Cite this article

Bogush, T.A., Tikhomirov, M.V., Dudko, E.A. et al. Quantitative immunofluorescence estimation of Pgp expression in human solid tumors by flow cytometry. Moscow Univ. Chem. Bull. 67, 142–148 (2012). https://doi.org/10.3103/S0027131412030054

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S0027131412030054

Keywords

Search

Navigation