Lectin microarray profiling of metastatic breast cancers

Abstract

Altered protein glycosylation compared with the disease-free state is a universal feature of cancer cells. It has long been established that distinct glycan structures are associated with specific forms of cancer, but far less is known about the complete array of glycans associated with certain tumors. The cancer glycome has great potential as a source of biomarkers, but progress in this field has been hindered by a lack of available techniques for the elucidation of disease-associated glycosylation. In the present study, lectin microarrays consisting of 45 lectins with different binding preferences covering N- and O-linked glycans were coupled with evanescent-field activated fluorescent detection in the glycomic analysis of primary breast tumors and the serum and urine of patients with metastatic breast cancer. A single 50 µm section of a primary breast tumor or <1 µL of breast cancer patient serum or urine was sufficient to detect glycosylation alterations associated with metastatic breast cancer, as inferred from lectin-binding patterns. The high-throughput, sensitive and relatively simple nature of the simultaneous analysis of N- and O-linked glycosylation following minimal sample preparation and without the need for protein deglycosylation makes the lectin microarray analysis described a valuable tool for discovery phase glycomic profiling.

Alteration of protein glycosylation in human hepatic stellate cells activated with transforming growth factor-β1

“…The most significant advantage of lectin microarrays over alternative methods of glycan profiling is that following minimal sample preparation the simultaneous quantitative analysis of N-and O-linked glycans on intact biological structures can be performed without the need for glycan release [26], which guarantees the real state of protein glycosylation in sample being correctly reflected. Recently, some improvements have been made on lectin microarray technique, i.e., building antibody-assisted lectin profiling or sandwich arrays [27,28], that is more conductive to attain specific signals corresponding to the target glycoprotein glycans at a sub-picomole level with the aid of specific antibodies.…”

Overexpression of α2,3sialyl T-antigen in breast cancer determined by miniaturized glycosyltransferase assays and confirmed using tissue microarray immunohistochemical analysis

“…Lectin based strategies have measured glycans in the serum of colon tumor patients [19], and in tumor, blood and urine samples of breast cancer patients [20]. Although such studies demonstrate differences between metastatic and non-metastatic breast cancer, the relatively broad binding specificity of lectins precludes the identification of unique changes in glycans or associated sugar linkages.…”

Serum paraoxonase 1 heteroplasmon, a fucosylated, and sialylated glycoprotein in distinguishing early hepatocellular carcinoma from liver cirrhosis patients

“…It is highly suitable for live cell or clinical fluid analysis and has the potential for high-throughput, low-cost, and fast discovery of glycan-related biomarkers [22,23]. Recently, several reports on lectin arrays have been published to screen glycan-related biomarkers of various diseases, such as metastatic breast cancer [24] and liver fibrosis [14]. Based on the lectin array profiling of early HCC, our study focuses on aberrant glycan structures of a particular protein which may have higher specificity as a biomarker for early HCC diagnosis.…”