Her2/neu extracellular domain shedding in uterine serous carcinoma

Implications for immunotherapy with trastuzumab

P. Todeschini, E. Cocco, S. Bellone, J. Varughese, Ken Yu Lin, L. Carrara, F. Guzzo, N. Buza, P. Hui, D. A. Silasi, E. Ratner, M. Azodi, P. E. Schwartz, T. J. Rutherford, S. Pecorelli, A. D. Santin

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Background: We evaluated shedding of epidermal growth factor type II receptor (Her2/neu) extracellular domain (ECD) in primary uterine serous carcinoma (USC) cell lines and in the serum of USC patients and its biological effects in experiments of trastuzumab-induced cytotoxicity in vitro. Methods: Her2/neu expression was evaluated by immunohistochemistry (IHC), real-time PCR and flow cytometry, while c-erbB2 gene amplification was assessed using fluorescent in situ hybridisation (FISH). Her2/neu ECD levels in the supernatants of USC cell lines and in the serum of 38 USC patients and 19 controls were tested using ELISA. The biologic effect of Her2/neu ECD on trastuzumab-induced antibody-dependent cell-mediated cytotoxicity (ADCC) was evaluated in 5-h chromium-release assays. Results: Five out of ten USC cell lines overexpressed Her2/neu by IHC and showed amplification of the c-erbB2 gene. High levels of Her2/neu ECD were found in supernatants of all FISH-positive tumours. In contrast, FISH-negative USC was negative for Her2/neu ECD shedding. Serum Her2/neu ECD levels in patients harbouring 3+Her2/neu tumours were higher than those found in healthy women (P=0.02) or USC patients with 2+ or 1+/negative Her2/neu expression (P=0.02). In cytotoxicity experiments, trastuzumab-mediated ADCC was significantly decreased by the addition of Her2/neu ECD-containing supernatants (P=0.01). Conclusion: FISH-positive c-erbB2 USC cell lines shed high levels of Her2/neu ECD. High levels of Her2/neu ECD in USC patients may reduce trastuzumab-mediated ADCC in vitro and potentially neutralise its therapeutic effect in vivo.

Original languageEnglish (US)
Pages (from-to)1176-1182
Number of pages7
JournalBritish Journal of Cancer
Volume105
Issue number8
DOIs
StatePublished - Oct 11 2011
Externally publishedYes

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Immunotherapy
Carcinoma
Antibody-Dependent Cell Cytotoxicity
Fluorescence In Situ Hybridization
erbB-2 Genes
Cell Line
Serum
Immunohistochemistry
Trastuzumab
Gene Amplification
Therapeutic Uses
Chromium
Epidermal Growth Factor Receptor
Real-Time Polymerase Chain Reaction
Neoplasms
Flow Cytometry
Enzyme-Linked Immunosorbent Assay

Keywords

  • Her2/neu
  • Her2/neu extracellular domain (ECD)
  • trastuzumab
  • uterine serous cancer

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Her2/neu extracellular domain shedding in uterine serous carcinoma : Implications for immunotherapy with trastuzumab. / Todeschini, P.; Cocco, E.; Bellone, S.; Varughese, J.; Lin, Ken Yu; Carrara, L.; Guzzo, F.; Buza, N.; Hui, P.; Silasi, D. A.; Ratner, E.; Azodi, M.; Schwartz, P. E.; Rutherford, T. J.; Pecorelli, S.; Santin, A. D.

In: British Journal of Cancer, Vol. 105, No. 8, 11.10.2011, p. 1176-1182.

Research output: Contribution to journalArticle

Todeschini, P, Cocco, E, Bellone, S, Varughese, J, Lin, KY, Carrara, L, Guzzo, F, Buza, N, Hui, P, Silasi, DA, Ratner, E, Azodi, M, Schwartz, PE, Rutherford, TJ, Pecorelli, S & Santin, AD 2011, 'Her2/neu extracellular domain shedding in uterine serous carcinoma: Implications for immunotherapy with trastuzumab', British Journal of Cancer, vol. 105, no. 8, pp. 1176-1182. https://doi.org/10.1038/bjc.2011.369
Todeschini, P. ; Cocco, E. ; Bellone, S. ; Varughese, J. ; Lin, Ken Yu ; Carrara, L. ; Guzzo, F. ; Buza, N. ; Hui, P. ; Silasi, D. A. ; Ratner, E. ; Azodi, M. ; Schwartz, P. E. ; Rutherford, T. J. ; Pecorelli, S. ; Santin, A. D. / Her2/neu extracellular domain shedding in uterine serous carcinoma : Implications for immunotherapy with trastuzumab. In: British Journal of Cancer. 2011 ; Vol. 105, No. 8. pp. 1176-1182.
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abstract = "Background: We evaluated shedding of epidermal growth factor type II receptor (Her2/neu) extracellular domain (ECD) in primary uterine serous carcinoma (USC) cell lines and in the serum of USC patients and its biological effects in experiments of trastuzumab-induced cytotoxicity in vitro. Methods: Her2/neu expression was evaluated by immunohistochemistry (IHC), real-time PCR and flow cytometry, while c-erbB2 gene amplification was assessed using fluorescent in situ hybridisation (FISH). Her2/neu ECD levels in the supernatants of USC cell lines and in the serum of 38 USC patients and 19 controls were tested using ELISA. The biologic effect of Her2/neu ECD on trastuzumab-induced antibody-dependent cell-mediated cytotoxicity (ADCC) was evaluated in 5-h chromium-release assays. Results: Five out of ten USC cell lines overexpressed Her2/neu by IHC and showed amplification of the c-erbB2 gene. High levels of Her2/neu ECD were found in supernatants of all FISH-positive tumours. In contrast, FISH-negative USC was negative for Her2/neu ECD shedding. Serum Her2/neu ECD levels in patients harbouring 3+Her2/neu tumours were higher than those found in healthy women (P=0.02) or USC patients with 2+ or 1+/negative Her2/neu expression (P=0.02). In cytotoxicity experiments, trastuzumab-mediated ADCC was significantly decreased by the addition of Her2/neu ECD-containing supernatants (P=0.01). Conclusion: FISH-positive c-erbB2 USC cell lines shed high levels of Her2/neu ECD. High levels of Her2/neu ECD in USC patients may reduce trastuzumab-mediated ADCC in vitro and potentially neutralise its therapeutic effect in vivo.",
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T1 - Her2/neu extracellular domain shedding in uterine serous carcinoma

T2 - Implications for immunotherapy with trastuzumab

AU - Todeschini, P.

AU - Cocco, E.

AU - Bellone, S.

AU - Varughese, J.

AU - Lin, Ken Yu

AU - Carrara, L.

AU - Guzzo, F.

AU - Buza, N.

AU - Hui, P.

AU - Silasi, D. A.

AU - Ratner, E.

AU - Azodi, M.

AU - Schwartz, P. E.

AU - Rutherford, T. J.

AU - Pecorelli, S.

AU - Santin, A. D.

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