title: Choline and phosphatidylcholine fluorescent derivatives localization in carcinoma cells studied by laser scanning confocal fluorescence microscopy.
abstract: In this study, we have shown the intracellular distribution of choline and phosphatidylcholine fluorescent derivatives in human breast carcinoma cells using confocal microscopy. The fluorescent choline derivatives ethanamimium 2-hydroxy-N,N-dimethyl-N-[2-N-(2,1,3-benzoxadiazol-4-amine,-N-methyl,-7- nitro)-ethyl] bromide (NBD-choline) and C(6)-NBD-phosphatidylcholine (C(6)-NBD-PC) were used in this work. NBD-choline was easily internalised into drug sensitive MCF-7 and in multidrug resistant MCF-7/DX cells. The probe was found to localise in the endoplasmic reticulum of sensitive cells and in the Golgi of multidrug resistant cells. In contrast, very low accumulation was found in normal MCF10A cells. For C(6)-NBD-PC, a similar pattern of localisation was found in tumour cells, but a significant uptake was also observed in normal cells. Unlike NBD-choline, C(6)-NBD-PC appears not to discriminate between normal and tumour cells. These results are consistent with previously published results showing higher levels of (11)C-choline uptake in malignant lesions seen with positron emission tomography (PET) in vivo imaging. Our results suggest that using NBD-choline and laser scanning confocal fluorescence microscopy (LSCFM) could be a useful tool to study choline metabolism in cancer cells and to consolidate PET imaging findings.
authors.affiliation: Department of Biotechnologies and Biosciences, University of Milano-Bicocca, Italy.
authors.forename: Anna M
authors.initials: AM
authors.lastname: Villa
authors.forename: Elvira
authors.initials: E
authors.lastname: Caporizzo
authors.forename: Antonio
authors.initials: A
authors.lastname: Papagni
authors.forename: Luciano
authors.initials: L
authors.lastname: Miozzo
authors.forename: Paola
authors.initials: P
authors.lastname: Del Buttero
authors.forename: Maria Donata
authors.initials: MD
authors.lastname: Grilli
authors.forename: Nadia
authors.initials: N
authors.lastname: Amboldi
authors.forename: Ferruccio
authors.initials: F
authors.lastname: Fazio
authors.forename: Silvia M
authors.initials: SM
authors.lastname: Doglia
authors.forename: Barbara
authors.initials: B
authors.lastname: Giglioni
mesh_terms: D001943:Breast Neoplasms / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D002794:Choline / Q000031:analogs & derivatives / Q000378:metabolism*; D005260:Female; D005453:Fluorescence; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D010713:Phosphatidylcholines / Q000032:analysis*; D049268:Positron-Emission Tomography; D014407:Tumor Cells, Cultured
chemical_list: D010713:Phosphatidylcholines; D002794:Choline