DBA Lectin Binds to Highly Proliferative Mouse Erythroleukemia Cells.
Fjalë kyçe
Abstrakt
OBJECTIVE
Hematopoietic malignancies lead to disease states involving abnormal proliferation of blood cells. Ki-67 and carboxyfluorescein succinimidyl ester (CFSE) are assays used to examine the proliferation status of cells but affect cell viability. In this study, we used lectins to bind to surfaces of proliferating cells with different phenotypes while preserving cell viability.
METHODS
The mouse lymphocyte Friend leukemia F5-5.F1 cell line was stained using biotin-conjugated lectins from Canavalia ensiformis (ConA), Dolichos biflorus (DBA), Erythrina cristagalli (ECA), Lens culinaris (LCA), Phaseolus vulgaris (PHA-E4), Arachis hypogaea (PNA), Ulex europaeus (UEA) and Triticum vulgaris (WGA) and sorted by fluorescence-activated cell sorting. Morphology, gene expression and proliferation assays were performed on sorted cells.
RESULTS
DBA, LCA and PHA-E4 probing sorted cells based on surface phenotype. Gene expression analysis showed that myelocytomatosis oncogene (Myc), cyclin D1 (Ccnd1), and cyclinD2 (Ccnd2) were more highly expressed in the DBA(High) fraction than DBA(Int) and DBA(Neg) fractions. Ki-67 expression and MTS assay correlated with the DBA-binding pattern, with DBA(High) reflecting the highest proliferative tendency.
CONCLUSIONS
Labeling with DBA allows selection of proliferating cells using flow cytometry.
