SIMULTANEOUS RADIOLABEL, GENETIC TAGGING AND PROLIFERATION ASSAYS TO STUDY THE ORGAN DISTRIBUTION AND FATE OF METASTATIC CELLS

  • Authors:
    • T FUJIMAKI
    • LM ELLIS
    • CD BUCANA
    • R RADINSKY
    • JE PRICE
    • IJ FIDLER
  • View Affiliations

  • Published online on: June 1, 1993     https://doi.org/10.3892/ijo.2.6.895
  • Pages: 895-901
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Abstract

We compared the suitability of 3 techniques to study tumor cell survival in the lungs of mice and proliferation into metastases. Genetic tagging of tumor cells with the bacterial beta-galactosidase marker gene lacZ, radiolabeling of tumor cells with [I-125]IdUrd, and S-phase labeling of cells with bromodeoxyuridine (BrdUrd) were used simultaneously to track the fate of highly metastatic K-1735 X-21 melanoma cells injected into syngeneic C3H/HeN mice. The melanoma cells were transfected with a plasmid containing lacZ and neomycin resistance genes. After growth in selective medium, the cells were incubated in medium containing [I-125]IdUrd and then injected i.v. into mice. Lungs isolated at various times after i.v. injection were processed for staining with X-gal, radioactive monitoring, and immunohistochemical staining with a monoclonal antibody against BrdUrd. At early time points, the presence of lacZ-positive cells directly correlated with radioactivity associated with viable cells. However, the expression of the beta-galactosidase was only stable for 1 week, and by 3 weeks after injection, large metastases contained only a few lacZ-positive cells. The combination of lacZ tagging with BrdUrd proliferation assay accurately identified dividing tumor cells in micrometastases. The simultaneous use of these 3 techniques allowed us to conclude that quantitative analysis of tumor cell survival is best accomplished by radioactive labeling of cells, whereas the use of lacZ-tagged cells allowed for studies of localization. Analysis of tumor cell proliferation requires the use of both lacZ tagging and immunohistochemistry using anti-BrdUrd antibodies. Since the process of metastasis consists of a series of distinct steps, each technique presents its own advantages and limitations.

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June 1993
Volume 2 Issue 6

Print ISSN: 1019-6439
Online ISSN:1791-2423

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APA
FUJIMAKI, T., ELLIS, L., BUCANA, C., RADINSKY, R., PRICE, J., & FIDLER, I. (1993). SIMULTANEOUS RADIOLABEL, GENETIC TAGGING AND PROLIFERATION ASSAYS TO STUDY THE ORGAN DISTRIBUTION AND FATE OF METASTATIC CELLS. International Journal of Oncology, 2, 895-901. https://doi.org/10.3892/ijo.2.6.895
MLA
FUJIMAKI, T., ELLIS, L., BUCANA, C., RADINSKY, R., PRICE, J., FIDLER, I."SIMULTANEOUS RADIOLABEL, GENETIC TAGGING AND PROLIFERATION ASSAYS TO STUDY THE ORGAN DISTRIBUTION AND FATE OF METASTATIC CELLS". International Journal of Oncology 2.6 (1993): 895-901.
Chicago
FUJIMAKI, T., ELLIS, L., BUCANA, C., RADINSKY, R., PRICE, J., FIDLER, I."SIMULTANEOUS RADIOLABEL, GENETIC TAGGING AND PROLIFERATION ASSAYS TO STUDY THE ORGAN DISTRIBUTION AND FATE OF METASTATIC CELLS". International Journal of Oncology 2, no. 6 (1993): 895-901. https://doi.org/10.3892/ijo.2.6.895