An in vivo tumor model expressing green fluorescent protein for the investigation of metastasis

  • Authors:
    • Oliver Thews
    • Christine Lambert
    • Debra K. Kelleher
    • Hans K. Biesalski
    • Peter Vaupel
    • Juergen Frank
  • View Affiliations

  • Published online on: September 1, 2005     https://doi.org/10.3892/ijo.27.3.705
  • Pages: 705-712
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Abstract

Although strong evidence is available suggesting that microenvironmental parameters play a role in lymphogenic or hematogenic metastasis, the underlying mechanisms are still unclear and further investigations of this topic are needed. For such a study however, an appropriate model of metastasis for in vivo analysis of this process would be required. An in vivo model of a solid tumor (rat DS sarcoma) has therefore been established to enable monitoring of the steps involved in tumor metastasis. Rat DS sarcoma cells were transfected with the pTracer-SV40 plasmid, containing the super-GFP and zeocin resistance genes. DS sarcoma cells showing high and stable expression of GFP (DSGFP cells) were selected by cell sorting and in vitro culturing with zeocin. To establish in vivo growth, DSGFP cells were subsequently injected intraperitoneally (i.p.) without additional selection by zeocin and GFP expression was monitored by flow cytometry. Using DSGFP ascites cells, solid tumors were implanted subcutaneously into the hind foot dorsum of rats. The expression of GFP was assayed by fluorescence microscopy. The detection of circulating DSGFP sarcoma cells in the blood was performed using the PCR technique. GFP expression in vitro was stable for more than 40 passages. Cell sorting, however, did not enable selection of a DSGFP cell population with a higher long-term stable GFP expression. After i.p. cell implantation, GFP expression in DSGFP ascites cells was maintained over at least 19 passages. Solid tumors implanted by injection of DSGFP ascites cells showed stable GFP expression. The growth rate of solid DSGFP sarcomas was slightly slower compared to that of non-transfected cell lines. The detection limit for circulating DS sarcoma cells in blood was 100 DSGFP cells/ml whole rat blood. Micrometastases in loco-regional lymph nodes, lung and liver were detectable by immunohistology and real-time PCR. This in vivo model showing stable expression of GFP could be useful for analyzing the mechanisms of metastasis, particularly where micrometastases or circulating tumor cells are to be identified.

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September 2005
Volume 27 Issue 3

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

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Spandidos Publications style
Thews O, Lambert C, Kelleher DK, Biesalski HK, Vaupel P and Frank J: An in vivo tumor model expressing green fluorescent protein for the investigation of metastasis. Int J Oncol 27: 705-712, 2005
APA
Thews, O., Lambert, C., Kelleher, D.K., Biesalski, H.K., Vaupel, P., & Frank, J. (2005). An in vivo tumor model expressing green fluorescent protein for the investigation of metastasis. International Journal of Oncology, 27, 705-712. https://doi.org/10.3892/ijo.27.3.705
MLA
Thews, O., Lambert, C., Kelleher, D. K., Biesalski, H. K., Vaupel, P., Frank, J."An in vivo tumor model expressing green fluorescent protein for the investigation of metastasis". International Journal of Oncology 27.3 (2005): 705-712.
Chicago
Thews, O., Lambert, C., Kelleher, D. K., Biesalski, H. K., Vaupel, P., Frank, J."An in vivo tumor model expressing green fluorescent protein for the investigation of metastasis". International Journal of Oncology 27, no. 3 (2005): 705-712. https://doi.org/10.3892/ijo.27.3.705