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. 2004 Sep 13;91(6):1195-9.
doi: 10.1038/sj.bjc.6602089.

Recombinant Humanised anti-HER2/neu Antibody (Herceptin) Induces Cellular Death of Glioblastomas

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Free PMC article

Recombinant Humanised anti-HER2/neu Antibody (Herceptin) Induces Cellular Death of Glioblastomas

J-F Mineo et al. Br J Cancer. .
Free PMC article

Abstract

Glioblastoma multiforme (GBM) remains the most devastating primary tumour in neuro-oncology. Targeting of the human epithelial receptor type 2 (HER2)-neu receptor by specific antibodies is a recent well-established therapy for breast tumours. Human epithelial receptor type 2/neu is a transmembrane tyrosine/kinase receptor that appears to be important for the regulation of cancer growth. Human epithelial receptor type 2/neu is not expressed in the adult central nervous system, but its expression increases with the degree of astrocytoma anaplasia. The specificity of HER2/neu for tumoral astrocytomas leads us to study in vitro treatment of GBM with anti-HER2/neu antibody. We used human GBM cell lines expressing HER2/neu (A172 express HER2/neu more than U251MG) or not (U87MG) and monoclonal humanised antibody against HER2/neu (Herceptin). Human epithelial receptor type 2/neu expression was measured by immunohistochemistry and flow cytometry. Direct antibody effect, complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity were evaluated by different cytometric assays. We have shown, for the first time, the ability of anti-HER2/neu antibodies to induce apoptosis and cellular-dependent cytotoxicity of HER2/neu-expressing GBM cell lines. The results decreased from A172 to U251 and were negative for U87MG, in accordance with the decreasing density of HER2/neu receptors.

Figures

Figure 1
Figure 1
Apoptosis induction: The A172 cell line expressing HER2/neu undergo apoptosis, following incubation with Herceptin®. Cells were incubated for 24 h with medium (A) or with 25 μg of Herceptin® (B). The cells staining with FITC-conjugated Annexin V and PI. Apoptotic cells were identified as Annexin V positive and PI negative. (C) Time course analysis of PS exposure after incubation of A172 cell line Herceptin® 25 μg ml−1. Only 24 h results are statistically significant (P<0.01).
Figure 2
Figure 2
Increasing apoptosis in glioblastomas cell lines expressing HER2/neu: Cells were incubated for 24 h with different concentrations of Herceptin®.
Figure 3
Figure 3
CD45 expression on the GBM cell lines and leucocytes: Cells were stained by FITC-conjugated mouse anti-human CD45 for 30 min at 4°C. Flow cytometry allowed us to differentiate the glioblastoma and effector cells.
Figure 4
Figure 4
Antibody-dependent cellular cytotoxicity of Herceptin® against A172 cell line. Target cells were incubated with effector cells (ratio targets/effectors 1/75) for 6 h with medium only (A) or with 10 μg ml−1 of Herceptin® (B). FITC labelled anti-human CD45 antibody then PI was added. Analysis was based on the negative A172 population CD45 determination. The dead cells were stained by PI (C).

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