Abstract
Purpose
Although a number of prognostic indicators have been developed, it is still difficult to predict the biological behaviour of all cancer types. 99mTc-(V)-DMSA (V DMSA) uptake and focal adhesion kinase (FAK) expression and activation level could be potential agents for this purpose. We hypothesised the existence of a correlation between V DMSA, whose uptake is linked to phosphate ions, essential compounds for tumour growth and cell proliferation, and the adhesion protein FAK, whose elevated expression and level of constitutive activation are implicated in cancer progression. The aim of this study was to assess the relationship between V DMSA incorporation rate and FAK expression and activation by phosphorylation on tyrosine 397 residue.
Methods
We determined V DMSA uptake in six different cancer cell lines and we measured FAK expression and activation by using Western Blotting analysis. Correlations with factors known to be associated with poor prognosis, such as invasive potential, resistance to chemotherapy and proliferation rate, were also investigated.
Results
The cell lines exhibited different V DMSA incorporation rates. In addition, these cells showed the same FAK expression, but various degrees of activation. A correlation was observed between V DMSA uptake and level of FAK phosphorylation and between V DMSA or constitutive FAK activation and proliferation rate. However, no correlation was shown between these parameters and the other factors tested, i.e. invasive potential and anticancer drug resistance.
Conclusion
The results of this in vitro study clearly demonstrate that phosphorylation of FAK, proliferation rate and V DMSA uptake are closely related. Because proliferation and a high level of constitutive FAK activation are linked to cancer progression, it can be assumed that in vivo V DMSA uptake reflects tumour aggressiveness.
Similar content being viewed by others
References
Schaller MD, Schaefer EM. Multiple stimuli induce tyrosine phosphorylation of the Crk-binding sites of paxillin. Biochem J 2001;360:57–66.
Lee LT, Huang YT, Hwang JJ, Lee AY, Ke FC, Huang CJ, et al. Transinactivation of the epidermal growth factor receptor tyrosine kinase and focal adhesion kinase phosphorylation by dietary flavonoids: effect on invasive potential of human carcinoma cells. Biochem Pharmacol 2004; 67:2103–14.
Wang JF, Park IW, Groopman JE. Stromal cell-derived factor-1alpha stimulates tyrosine phosphorylation of multiple focal adhesion proteins and induces migration of hematopoietic progenitor cells: roles of phosphoinositide-3 kinase and protein kinase C. Blood 2000;95:2505–13.
Sakurai S, Sonoda Y, Koguchi E, Shinoura N, Hamada H, Kasahara T. Mutated focal adhesion kinase induces apoptosis in a human glioma cell line, T98G. Biochem Biophys Res Commun 2002;293:174–81.
Sieg DJ, Hauck CR, Schlaepfer DD. Required role of focal adhesion kinase (FAK) for integrin-stimulated cell migration. J Cell Sci 1999;112:2677–91.
Cornillon J, Campos L, Guyotat D. Focal adhesion kinase (FAK), une protéine aux fonctions multiples. Med Sci (Paris) 2003;19:743–52.
Gabarra-Niecko V, Schaller MD, Dunty JM. FAK regulates biological processes important for the pathogenesis of cancer. Cancer Metastasis Rev 2003;22:359–74.
Duxbury MS, Ito H, Benoit E, Zinner MJ, Ashley SW, Whang EE. RNA interference targeting focal adhesion kinase enhances pancreatic adenocarcinoma gemcitabine chemosensitivity. Biochem Biophys Res Commun 2003;311:786–92.
Sonoda Y, Matsumoto Y, Funakoshi M, Yamamoto D, Hanks SK, Kasahara T. Anti-apoptotic role of focal adhesion kinase (FAK). Induction of inhibitor-of-apoptosis proteins and apoptosis suppression by the overexpression of FAK in a human leukemic cell line, HL-60. J Biol Chem 2000;275:16309–15.
Papantoniou VJ, Souvatzoglou MA, Valotassiou VJ, Louvrou AN, Ambela C, Koutsikos J, et al. Relationship of cell proliferation (Ki-67) to 99mTc-(V)DMSA uptake in breast cancer. Breast Cancer Res 2004;6:R56–R62.
Papantoniou V, Christodoulidou J, Papadaki E, Valotassiou V, Souvatzoglou M, Louvrou A, et al. Uptake and washout of 99mTcV-dimercaptosuccinic acid and 99mTc-sestamibi in the assessment of histological type and grade in breast cancer. Nucl Med Commun 2002;23:461–7.
Hirano T, Otake H, Kazama K, Wakabayashi K, Zama A, Shibasaki T, et al. Technetium-99m(V)-DMSA and thallium-201 in brain tumor imaging: correlation with histology and malignant grade. J Nucl Med 1997; 38:1741–9.
Hirano T, Otake H, Yoshida I, Endo K. Primary lung cancer SPECT imaging with pentavalent technetium-99m-DMSA. J Nucl Med 1995;36:202–7.
Atasever T, Gundogdu C, Vural G, Kapucu LO, Karalezli A, Unlu M. Evaluation of pentavalent Tc-99m DMSA scintigraphy in small cell and nonsmall cell lung cancers. Nuklearmedizin 1997;36:223–7.
Lam AS, Kettle AG, O’Doherty MJ, Coakley AJ, Barrington SF, Blower PJ. Pentavalent 99Tcm-DMSA imaging in patients with bone metastases. Nucl Med Commun 1997;18:907–14.
Babbar A, Kashyap R, Chauhan UP. A convenient method for the preparation of 99mTc-labelled pentavalent DMSA and its evaluation as a tumour imaging agent. J Nucl Biol Med 1991;35:100–4.
Kashyap R, Babbar A, Sahai I, Prakash R, Soni NL, Chauhan UP. Tc-99 m(V) DMSA imaging. A new approach to studying metastases from breast carcinoma. Clin Nucl Med 1992;17:119–22.
Kiratli H, Kiratli PO, Ercan MT. Scintigraphic evaluation of tumors metastatic to the choroid using technetium-99m(V)-dimercaptosuccinic acid. Jpn J Ophthalmol 1998;42:60–5.
Denoyer D, Perek N, Le Jeune N, Frere D, Dubois F. Evidence that 99mTc-(V)-DMSA uptake is mediated by NaPi cotransporter type III in tumour cell lines. Eur J Nucl Med Mol Imaging 2004;31:77–84.
Beauchesne P, Bertrand S, MJ Ng, Christianson T, Dore JF, Mornex F, et al. Etoposide sensitivity of radioresistant human glioma cell lines. Cancer Chemother Pharmacol 1998;41:93–7.
Ke LD, Shi YX, Im SA, Chen X, Yung WK. The relevance of cell proliferation, vascular endothelial growth factor, and basic fibroblast growth factor production to angiogenesis and tumorigenicity in human glioma cell lines. Clin Cancer Res 2000;6:2562–72.
Hirano T, Otake H, Shibasaki T, Tamura M, Endo K. Differentiating histologic malignancy of primary brain tumors: pentavalent technetium-99m-DMSA. J Nucl Med 1997;38:20–6.
Kahana O, Micksche M, Witz IP, Yron I. The focal adhesion kinase (P125FAK) is constitutively active in human malignant melanoma. Oncogene 2002;21:3969–77.
Ala-Aho R, Johansson N, Baker AH, Kahari VM. Expression of collagenase-3 (MMP-13) enhances invasion of human fibrosarcoma HT-1080 cells. Int J Cancer 2002;97:283–9.
Sato J, Sata M, Nakamura H, Inoue S, Wada T, Takabatake N, et al. Role of thymidine phosphorylase on invasiveness and metastasis in lung adenocarcinoma. Int J Cancer 2003;106:863–70.
Ruohola JK, Viitanen TP, Valve EM, Seppanen JA, Loponen NT, Keskitalo JJ, et al. Enhanced invasion and tumor growth of fibroblast growth factor 8b-overexpressing MCF-7 human breast cancer cells. Cancer Res 2001;61:4229–37.
Owens LV, Xu L, Craven RJ, Dent GA, Weiner TM, Kornberg L, et al. Overexpression of the focal adhesion kinase (p125FAK) in invasive human tumors. Cancer Res 1995;55:2752–5.
Aloj L, Zannetti A, Caracó C, Del Vecchio S, Salvatore M. Bcl-2 overexpression prevents 99mTc-MIBI uptake in breast cancer cell lines. Eur J Nucl Med Mol Imaging 2004;31:521–7.
Cory S, Adams JM. The Bcl-2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2002;2:647–56.
Denoyer D, Perek N, Jeune NL, Frere D, Sabido O, Clotagatide A, et al. In vitro and in vivo evaluation of the influence of type III NaPi co-transporter activity during apoptosis on 99mTc-(V)DMSA uptake in the human leukaemic cell line U937. Eur J Nucl Med Mol Imaging 2004;31:1421–7.
Acknowledgements
This work was supported by grants from the Ligue Nationale Contre le Cancer-Comité Départemental de la Loire. We thank Mr. Saul for his help in translation of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Denoyer, D., Perek, N., Le Jeune, N. et al. Correlation between 99mTc-(V)-DMSA uptake and constitutive level of phosphorylated focal adhesion kinase in an in vitro model of cancer cell lines. Eur J Nucl Med Mol Imaging 32, 820–827 (2005). https://doi.org/10.1007/s00259-005-1773-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00259-005-1773-4