Heparan sulfate mediates trastuzumab effect in breast cancer cells

BMC Cancer

Eloah Rabello Suarez³  Edgar Julian Paredes-Gamero²  Auro Del Giglio¹  Ivarne Luis dos Santos Tersariol²  Helena Bonciani Nader²  Maria Aparecida Silva Pinhal³ 

[1] Department of Oncology, Faculdade de Medicina do ABC, Avenida Príncipe de Gales, 821, Vila Príncipe de Gales, 09060-650, Santo André, SP, Brazil

[2] Department of Biochemistry, Universidade Federal de São Paulo, Rua Três de Maio, 100, Vila Clementino, 04044-020, São Paulo, SP, Brazil

[3] Department of Biochemistry, Faculdade de Medicina do ABC, Avenida Príncipe de Gales, 821, Vila Príncipe de Gales, 09060-650, Santo André, SP, Brazil

关键词: Breast cancer resistance;    Glycoaminoglycans;    Proteoglycans;    Dermatan sulfate;    Heparan sulfate;    Heparanase;    HER2;   

Others  :  1079547
DOI  :  10.1186/1471-2407-13-444

 received in 2013-07-23, accepted in 2013-09-26,  发布年份 2013

【 摘 要 】

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

Trastuzumab action is dependent on the availability of heparan sulfate on the surface of breast cancer cells. Furthermore, our data suggest that high levels of heparan sulfate shed to the medium are able to capture trastuzumab, blocking the antibody action mediated by HER2. In addition to HER2 levels, heparan sulfate synthesis and shedding determine breast cancer cell susceptibility to trastuzumab.

【 授权许可】

   
2013 Suarez et al.; licensee BioMed Central Ltd.

【 图 表 】

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