Abstract | BACKGROUND:
Trastuzumab is an antibody widely used in the treatment of breast cancer cases that test positive for the human epidermal growth factor receptor 2 (HER2). Many patients, however, become resistant to this antibody, whose resistance has become a major focus in breast cancer research. But despite this interest, there are still no reliable markers that can be used to identify resistant patients. A possible role of several extracellular matrix (ECM) components-- heparan sulfate (HS), Syn-1(Syndecan-1) and heparanase (HPSE1)--in light of the influence of ECM alterations on the action of several compounds on the cells and cancer development, was therefore investigated in breast cancer cell resistance to trastuzumab. METHODS: The cDNA of the enzyme responsible for cleaving HS chains from proteoglycans, HPSE1, was cloned in the pEGFP-N1 plasmid and transfected into a breast cancer cell lineage. We evaluated cell viability after trastuzumab treatment using different breast cancer cell lines. Trastuzumab and HS interaction was investigated by confocal microscopy and Fluorescence Resonance Energy Transfer (FRET). The profile of sulfated glycosaminoglycans was also investigated by [35S]- sulfate incorporation. Quantitative RT-PCR and immunofluorescence were used to evaluate HPSE1, HER2 and Syn-1 mRNA expression. HPSE1 enzymatic activity was performed using biotinylated heparan sulfate. RESULTS:
Breast cancer cell lines responsive to trastuzumab present higher amounts of HER2, Syn-1 and HS on the cell surface, but lower levels of secreted HS. Trastuzumab and HS interaction was proven by FRET analysis. The addition of anti-HS to the cells or heparin to the culture medium induced resistance to trastuzumab in breast cancer cells previously sensitive to this monoclonal antibody. Breast cancer cells transfected with HPSE1 became resistant to trastuzumab, showing lower levels of HER2, Syn-1 and HS on the cell surface. In addition, HS shedding was increased significantly in these resistant cells. CONCLUSION:
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Authors | Eloah Rabello Suarez, Edgar Julian Paredes-Gamero, Auro Del Giglio, Ivarne Luis dos Santos Tersariol, Helena Bonciani Nader, Maria Aparecida Silva Pinhal |
Journal | BMC cancer
(BMC Cancer)
Vol. 13
Pg. 444
(Oct 01 2013)
ISSN: 1471-2407 [Electronic] England |
PMID | 24083474
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Monoclonal, Humanized
- Antineoplastic Agents
- Glycosaminoglycans
- Syndecan-1
- A73025
- Heparitin Sulfate
- Receptor, ErbB-2
- heparanase
- Glucuronidase
- Trastuzumab
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Topics |
- Antibodies, Monoclonal, Humanized
(metabolism, pharmacology)
- Antineoplastic Agents
(metabolism, pharmacology)
- Breast Neoplasms
(genetics, metabolism)
- Cell Line, Tumor
- Cell Membrane
(metabolism)
- Cell Survival
(drug effects)
- Drug Resistance, Neoplasm
- Female
- Gene Expression Regulation, Neoplastic
- Glucuronidase
(genetics, metabolism)
- Glycosaminoglycans
(metabolism)
- Heparitin Sulfate
(metabolism)
- Humans
- MCF-7 Cells
- Protein Binding
- Protein Transport
- Receptor, ErbB-2
(antagonists & inhibitors, genetics, metabolism)
- Syndecan-1
(genetics, metabolism)
- Trastuzumab
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