Chemiluminescent Detection of Carbohydrates in the Tumoral Breast Diseases

Abstract

Nowadays, there is an increase of investigations into the fibroadenoma, mainly because some studies have shown that the occurrence of fibroadenoma is linked to an increased risk of developing breast carcinoma. Currently, the chemiluminescence biomarkers are applied for validation methods and screening. Here, a lectin chemiluminescence is proposed as new histochemistry method to identify carbohydrates in mammary tumoral tissues. The lectins concanavalin A (Con A) and peanut agglutinin (PNA) conjugated to acridinium ester were used to characterize the glycocode of breast tissues: normal, fibroadenoma, and invasive duct carcinoma (IDC). The lectin chemiluminescence expressed in relative light units (RLU) was higher in fibroadenoma and IDC than in normal tissue for both lectins tested. The relationship RLU emission versus tissue area described a linear and hyperbolic curve for IDC and fibroadenoma, respectively, using Con A whereas hyperbolic curves for both transformed tissues using PNA. RLU was abolished by inhibiting the interaction between tissues and lectins using their specific carbohydrates: methyl-α-D: -mannoside (Con A) and galactose (PNA). The intrinsic fluorescence emission did not change with combination of the lectins (Con A/PNA) to the acridinium ester for hydrophobic residues. These results represent the lectin chemiluminescence as an alternative of histochemistry method for tumoral diagnosis in the breast.

Glycophenotype Evaluation in Cutaneous Tumors Using Lectins Labeled with Acridinium Ester

“…Lectins conjugated to acridinium ester (Con A-AE, PNA-AE, UEA-AE, and MAA-AE) were collected by Sephadex G-25 chromatography and their profiles were identical to those previously reported [15]. The superposition of the protein and chemiluminescence peaks indicated that all studied lectins were labeled with the AE.…”

“…It is a promising and effective technique for quantitative analysis of proteins, nucleic acids, and carbohydrates [13–15]. Therefore, this approach combined with lectin histochemistry could be valuable to reduce the subjective evaluation.…”

Significance of immunohistochemistry in breast cancer

“…Due to the intrinsic subjectivity of the results of the two techniques (IHC and FISH), the identification of new methods with less subjectivity is necessary. Recently, the chemiluminescent technique has been used as a quantitative assay [39,40] . It has many benefits such as high sensitivity and accuracy, stability of reagents, easy-to-use protocols, non-photodegradable products, and a good correlation between IHC and immunohistochemiluminescence results.…”

Evaluation of glycophenotype in breast cancer by quantum dot-lectin histochemistry

“…16 On the other hand, Brustein et al, using Con A and peanut agglutinin lectins conjugated to acridinium ester, found that chemiluminescence was higher in fibroadenoma than in invasive ductal carcinoma for both lectins tested. 29 This shows that, in addition to the method of labeling or detection used, pretreatment of tissues, such as formalin-fixing and paraffin-embedding, plays an important role in the intensity …”