DBA lectin binds to highly proliferative mouse erythroleukemia cells

Anthony Swain, Kasem Kulkeaw, Yuka Tanaka, Yoichi Nakanishi, Senji Shirasawa, Daisuke Sugiyama

Research output: Contribution to journalArticlepeer-review

Abstract

Background/Aim: 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. Materials and 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 DBAHigh fraction than DBAInt and DBANeg fractions. Ki-67 expression and MTS assay correlated with the DBAbinding pattern, with DBAHigh reflecting the highest proliferative tendency. Conclusion: Labeling with DBA allows selection of proliferating cells using flow cytometry.

Original languageEnglish
Pages (from-to)3625-3633
Number of pages9
JournalAnticancer research
Volume36
Issue number7
Publication statusPublished - 2016

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

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