Glycophenotype Evaluation in Cutaneous Tumors Using Lectins Labeled with Acridinium Ester

Abstract

Background. Tumor cells show alterations in their glycosylation patterns when compared to normal cells. Lectins can be used to evaluate these glycocode changes. Chemiluminescence assay is an effective technique for quantitative analysis of proteins, nucleic acids, and carbohydrates due to its high sensitivity, specificity, and rapid testing. Objective. To use histochemiluminescence based on lectin conjugated to acridinium ester (AE) for the investigation of glycophenotype changes in cutaneous tumors. Methods. Concanavalin A (Con A), Peanut agglutinin (PNA), Ulex europaeus agglutinin-I (UEA-I), and Maackia amurensis agglutinin (MAA) were conjugated to acridinium ester. Biopsies of cutaneous tumors and normal skin were incubated with the lectins-AE, and chemiluminescence was quantified and expressed as Relative Light Units (RLU). Results. Actinic keratosis (AK), keratoacanthoma (KA), squamous cell carcinoma (SCC), and basal cell carcinoma (BCC) showed lower expression of α-D-glucose/mannose and α-L-fucose residues compared to normal tissue. Cutaneous tumors displayed higher expression of Gal-β(1-3)-GalNAc residues than normal tissue. AK and SCC exhibited higher expression of Neu5Ac-α(2,3)Gal residues than normal epidermis. KA and BCC showed equivalent RLU values compared to normal tissue. Conclusions. Lectin histochemiluminescence allowed quantitative assessment of the carbohydrate expression in cutaneous tissues, contributing to eliminate the subjectivity of conventional techniques used in the histopathological diagnosis.

Dimethyl‐2‐[(acridin‐9‐yl)methylidene]‐malonate as fluorescent probe for histochemical analysis

“…Aberrant glycosylation in glycoproteins and glycolipid is a common phenomenon associated with malignant transformation, invasion, and metastasis (Ghazarian, Idoni, & Oppenheimer, 2010). Lectins have been intensely used as histochemistry probes for cell surface saccharide profiling in normal and transformed tissues (Beltrão et al, 2003;Brustein et al, 2012;Campos, Cavalcanti, Lima-Filho, Carvalho, & Beltrão, 2006;Lima et al, 2013;Silva et al, 2014). Here, the glucose/mannose profile of breast tissues was detected using Con A conjugated to dimethyl 2-[(acridin-9-yl)methylidene]-malonate (LPSF/IP-81), an acridine derivative.…”

“…For instance, FITC has been conjugated to lectin for recognizing pterygium remnant after surgical excision (Díaz‐González et al, ). In our lab, lectins conjugated to different kinds of labels have been employed to analyzing the cancer glycophenotype in histochemistry (Brustein et al, ; Lima et al, ; Melo‐Júnior, Araújo‐Filho, Patú, Beltrão, & Carvalho, ; Santos et al, ; Silva et al, ). Lectins are structurally diverse carbohydrate‐binding proteins of non‐immune origin that agglutinate cells and recognize carbohydrates in oligosaccharides and glycoconjugates (Sharon, ).…”

“…In our lab, lectins conjugated to different kinds of labels have been employed to analyzing the cancer glycophenotype in histochemistry (Brustein et al, 2012;Lima et al, 2013;Melo-J unior, Ara ujo-Filho, Pat u, Beltrão, & Carvalho, 2006;Santos et al, 2006;Silva et al, 2014).…”

Cadherins and their Role in Malignant Transformation: Implications for Skin Cancer Progression

“…For example, AK, KA, SCC and BCC show lower expression of high mannose-type and/or hybrid-type N-glycans as well as fucose α1,2-linked to galactose residue (H antigen) compared to normal tissue. However, cutaneous tumours (SCC, BCC, invasive melanoma) display higher expression of truncated mucin-type O-glycan, that is, T antigen (Galβ1-3Gal-NAc residue) than normal tissue [111][112][113]. Mannose-type and/or hybrid-type N-glycans and T antigen can be used as markers for the distinction between BCC and TE [111].…”

“…Mannose-type and/or hybrid-type N-glycans and T antigen can be used as markers for the distinction between BCC and TE [111]. Higherexpression levels of β-galactoside α-2,3 sialyltransferase (ST3Gal I) and higher cell-surface reactivity with Maackia amurensis agglutinin (MAA), which recognizes sialic acid α2,3-linked to Gal residue, allow to distinguish AK and SCC from KA, BCC and normal epidermis [113,114]. MCCs do not show expression of neither α2,3-linked sialic acid nor H antigen [115].…”