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Effective treatment of chemoresistant breast cancer in vitro and in vivo by a factor VII-targeted photodynamic therapy.

Duanmu J, Cheng J, Xu J, Booth CJ, Hu Z - Br. J. Cancer (2011)

Bottom Line: The efficacy of fVII-tPDT was tested in vitro and in vivo and was compared with non-targeted PDT for treatment of chemoresistant breast cancer.Moreover, fVII-tPDT was effective and safe for the treatment of chemoresistant breast tumours in the nude mouse model.We conclude that fVII-tPDT is effective and safe for the treatment of chemoresistant breast cancer, presumably by simultaneously targeting both the tumour neovasculature and chemoresistant cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, 310 Cedar Street, New Haven, CT 06520, USA.

ABSTRACT

Background: The purpose of this study was to test a novel, dual tumour vascular endothelial cell (VEC)- and tumour cell-targeting factor VII-targeted Sn(IV) chlorin e6 photodynamic therapy (fVII-tPDT) by targeting a receptor tissue factor (TF) as an alternative treatment for chemoresistant breast cancer using a multidrug resistant (MDR) breast cancer line MCF-7/MDR.

Methods: The TF expression by the MCF-7/MDR breast cancer cells and tumour VECs in MCF-7/MDR tumours from mice was determined separately by flow cytometry and immunohistochemistry using anti-human or anti-murine TF antibodies. The efficacy of fVII-tPDT was tested in vitro and in vivo and was compared with non-targeted PDT for treatment of chemoresistant breast cancer. The in vitro efficacy was determined by a non-clonogenic assay using crystal violet staining for monolayers, and apoptosis and necrosis were assayed to elucidate the underlying mechanisms. The in vivo efficacy of fVII-tPDT was determined in a nude mouse model of subcutaneous MCF-7/MDR tumour xenograft by measuring tumour volume.

Results: To our knowledge, this is the first presentation showing that TF was expressed on tumour VECs in chemoresistant breast tumours from mice. The in vitro efficacy of fVII-tPDT was 12-fold stronger than that of ntPDT for MCF-7/MDR cancer cells, and the mechanism of action involved induction of apoptosis and necrosis. Moreover, fVII-tPDT was effective and safe for the treatment of chemoresistant breast tumours in the nude mouse model.

Conclusions: We conclude that fVII-tPDT is effective and safe for the treatment of chemoresistant breast cancer, presumably by simultaneously targeting both the tumour neovasculature and chemoresistant cancer cells. Thus, this dual-targeting fVII-tPDT could also have therapeutic potential for the treatment of other chemoresistant cancers.

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Factor VII-targeted photodynamic therapy (fVII-tPDT) induces significantly stronger levels of apoptosis and necrosis in MCF-7/MDR cancer cells compared with ntPDT. Necrosis (cytotoxicity) (A) and apoptosis (B) were detected in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. (C) Apoptosis and necrosis were visualised by immunofluorescence staining in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. Note that the fVII-tPDT-treated MCF-7/MDR cells were positively stained for necrosis (red in the nuclei) and apoptosis (green), whereas the ntPDT using unconjugated SnCe6 (2 μ, 36 J cm–2)-treated cells and the untreated control cells did not show any staining for apoptosis and necrosis. Note that the fVII-tPDT-treated MCF-7/MDR cells were clearly damaged, whereas the ntPDT-treated and untreated control cells were intact (phase contrast). Original magnification × 400.
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fig3: Factor VII-targeted photodynamic therapy (fVII-tPDT) induces significantly stronger levels of apoptosis and necrosis in MCF-7/MDR cancer cells compared with ntPDT. Necrosis (cytotoxicity) (A) and apoptosis (B) were detected in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. (C) Apoptosis and necrosis were visualised by immunofluorescence staining in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. Note that the fVII-tPDT-treated MCF-7/MDR cells were positively stained for necrosis (red in the nuclei) and apoptosis (green), whereas the ntPDT using unconjugated SnCe6 (2 μ, 36 J cm–2)-treated cells and the untreated control cells did not show any staining for apoptosis and necrosis. Note that the fVII-tPDT-treated MCF-7/MDR cells were clearly damaged, whereas the ntPDT-treated and untreated control cells were intact (phase contrast). Original magnification × 400.

Mentions: From the same experiments as in Figure 2C, we photographed the cancer cells before they were fixed for crystal violet staining. The fVII-tPDT-treated MCF-7/MDR cancer cells became debris even by laser fluence at 10.8 J cm–2, and none of the cancer cells remained intact after being treated with 43.2 J cm–2 fVII-tPDT (data not shown). In contrast, no cellular damage was observed in the untreated cells or any of the ntPDT-treated cells, even by the highest fluence (43.2 J cm–2; data not shown). Similar observations on cellular morphology are shown in Figure 3 below. We conclude that fVII targeting enhances the effect of SnCe6 PDT, and fVII-tPDT is effective in eradicating MCF-7/MDR cancer cells in vitro.


Effective treatment of chemoresistant breast cancer in vitro and in vivo by a factor VII-targeted photodynamic therapy.

Duanmu J, Cheng J, Xu J, Booth CJ, Hu Z - Br. J. Cancer (2011)

Factor VII-targeted photodynamic therapy (fVII-tPDT) induces significantly stronger levels of apoptosis and necrosis in MCF-7/MDR cancer cells compared with ntPDT. Necrosis (cytotoxicity) (A) and apoptosis (B) were detected in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. (C) Apoptosis and necrosis were visualised by immunofluorescence staining in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. Note that the fVII-tPDT-treated MCF-7/MDR cells were positively stained for necrosis (red in the nuclei) and apoptosis (green), whereas the ntPDT using unconjugated SnCe6 (2 μ, 36 J cm–2)-treated cells and the untreated control cells did not show any staining for apoptosis and necrosis. Note that the fVII-tPDT-treated MCF-7/MDR cells were clearly damaged, whereas the ntPDT-treated and untreated control cells were intact (phase contrast). Original magnification × 400.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3101942&req=5

fig3: Factor VII-targeted photodynamic therapy (fVII-tPDT) induces significantly stronger levels of apoptosis and necrosis in MCF-7/MDR cancer cells compared with ntPDT. Necrosis (cytotoxicity) (A) and apoptosis (B) were detected in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. (C) Apoptosis and necrosis were visualised by immunofluorescence staining in fVII-tPDT (2 μ SnCe6 and 36 J cm–2)-treated MCF-7/MDR cells. Note that the fVII-tPDT-treated MCF-7/MDR cells were positively stained for necrosis (red in the nuclei) and apoptosis (green), whereas the ntPDT using unconjugated SnCe6 (2 μ, 36 J cm–2)-treated cells and the untreated control cells did not show any staining for apoptosis and necrosis. Note that the fVII-tPDT-treated MCF-7/MDR cells were clearly damaged, whereas the ntPDT-treated and untreated control cells were intact (phase contrast). Original magnification × 400.
Mentions: From the same experiments as in Figure 2C, we photographed the cancer cells before they were fixed for crystal violet staining. The fVII-tPDT-treated MCF-7/MDR cancer cells became debris even by laser fluence at 10.8 J cm–2, and none of the cancer cells remained intact after being treated with 43.2 J cm–2 fVII-tPDT (data not shown). In contrast, no cellular damage was observed in the untreated cells or any of the ntPDT-treated cells, even by the highest fluence (43.2 J cm–2; data not shown). Similar observations on cellular morphology are shown in Figure 3 below. We conclude that fVII targeting enhances the effect of SnCe6 PDT, and fVII-tPDT is effective in eradicating MCF-7/MDR cancer cells in vitro.

Bottom Line: The efficacy of fVII-tPDT was tested in vitro and in vivo and was compared with non-targeted PDT for treatment of chemoresistant breast cancer.Moreover, fVII-tPDT was effective and safe for the treatment of chemoresistant breast tumours in the nude mouse model.We conclude that fVII-tPDT is effective and safe for the treatment of chemoresistant breast cancer, presumably by simultaneously targeting both the tumour neovasculature and chemoresistant cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, 310 Cedar Street, New Haven, CT 06520, USA.

ABSTRACT

Background: The purpose of this study was to test a novel, dual tumour vascular endothelial cell (VEC)- and tumour cell-targeting factor VII-targeted Sn(IV) chlorin e6 photodynamic therapy (fVII-tPDT) by targeting a receptor tissue factor (TF) as an alternative treatment for chemoresistant breast cancer using a multidrug resistant (MDR) breast cancer line MCF-7/MDR.

Methods: The TF expression by the MCF-7/MDR breast cancer cells and tumour VECs in MCF-7/MDR tumours from mice was determined separately by flow cytometry and immunohistochemistry using anti-human or anti-murine TF antibodies. The efficacy of fVII-tPDT was tested in vitro and in vivo and was compared with non-targeted PDT for treatment of chemoresistant breast cancer. The in vitro efficacy was determined by a non-clonogenic assay using crystal violet staining for monolayers, and apoptosis and necrosis were assayed to elucidate the underlying mechanisms. The in vivo efficacy of fVII-tPDT was determined in a nude mouse model of subcutaneous MCF-7/MDR tumour xenograft by measuring tumour volume.

Results: To our knowledge, this is the first presentation showing that TF was expressed on tumour VECs in chemoresistant breast tumours from mice. The in vitro efficacy of fVII-tPDT was 12-fold stronger than that of ntPDT for MCF-7/MDR cancer cells, and the mechanism of action involved induction of apoptosis and necrosis. Moreover, fVII-tPDT was effective and safe for the treatment of chemoresistant breast tumours in the nude mouse model.

Conclusions: We conclude that fVII-tPDT is effective and safe for the treatment of chemoresistant breast cancer, presumably by simultaneously targeting both the tumour neovasculature and chemoresistant cancer cells. Thus, this dual-targeting fVII-tPDT could also have therapeutic potential for the treatment of other chemoresistant cancers.

Show MeSH
Related in: MedlinePlus