NCAM polysialylation during adherence transitions: Live cell monitoring using an antibody-mimetic EGFP-endosialidase and the viability dye DRAQ7
作者： Paul J. Smith , Emeline Furon , Marie Wiltshire , Sally Chappell , Laurence H. Patterson
DOI: 10.1002/CYTO.A.22306
关键词:
摘要: Polysialylation of neural cell adhesion molecule (NCAM) in small-cell lung cancer (SCLC) is thought to regulate NCAM-mediated cell–surface interactions, imparting antiadhesive properties cells. However, SCLC cells culture demonstrate anchorage-independent growth and spontaneously generate adherent forms. Here, the ability polySia-NCAM influence proliferation adherence unclear. We analyzed live expression by flow cytometry, using novel combination a polySia antibody-mimetic eGFP-tagged endosialidase viability dye DRAQ7. Enrichment for ( 150 population doublings) resolved with stable re-expression increased rates both vitro vivo. Endoneuraminidase removal from re-expressing showed that rapid extracellular matrix components was functionally independent polySia. PolySia not altered when isolated forms underwent enforced cell–cell contact three-dimensional culture. Coculture variants modulated overall profiles indicating an microcommunity composition substrate potential. conclude obligatory linkage between potential rejected suggest degree homeostasis operates polysialylation within heterogeneous populations. The findings new model progression while application profiling could be used assess polySia-NCAM-targeted therapies. © 2013 International Society Advancement Cytometry
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