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Choline and phosphatidylcholine fluorescent derivatives localization in carcinoma cells studied by laser scanning confocal fluorescence microscopy. (Articolo in rivista)

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Choline and phosphatidylcholine fluorescent derivatives localization in carcinoma cells studied by laser scanning confocal fluorescence microscopy. (Articolo in rivista) (literal)

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Villa A.M. 1,2, Caporizzo E. 3, Papagni A. 2,4, Miozzo L. 2,4, Buttero P.D. 5, Grilli M.D. 5, Amboldi N. 6, Fazio F. 3, Doglia S.M. 1,2 , Giglioni B. 3 (2005)
Choline and phosphatidylcholine fluorescent derivatives localization in carcinoma cells studied by laser scanning confocal fluorescence microscopy.
in European journal of cancer (1990)
(literal)

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Villa A.M. 1,2, Caporizzo E. 3, Papagni A. 2,4, Miozzo L. 2,4, Buttero P.D. 5, Grilli M.D. 5, Amboldi N. 6, Fazio F. 3, Doglia S.M. 1,2 , Giglioni B. 3 (literal)

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The newly synthesised fluorescent NBD-choline, designed to probe choline uptake and distribution in living cells, has been successfully used in this LSCFM study to visualise the intracellular compartments involved in choline metabolism and choline-phospholipid biosynthesis in human breast tumour and normal cell lines. In particular, NBD-choline was found to predominantly localise in the ER of drug-sensitive MCF-7 cells and in the Golgi apparatus of multidrug resistant MCF-7/DX cells. This difference in NBD-choline distribution between drug-sensitive and drug resistant cell lines suggests for the first time that MDR cells have changed choline metabolic pathways. NBD-choline enters almost exclusively into cancer cells and not in normal cells, a result that confirms in vivo imaging by 11 C-choline-PET of several human tumours. The results of this study have shown the specificity of choline uptake by cancer cells and highlight the possibility of designing new choline derivatives that can be specifically targeted to tumour cells for anticancer activity. (literal)

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1. Department of Biotechnologies and Biosciences, University of Milano-Bicocca, Milan, Italy 2. INFM, UdR Milano-Bicocca, Milan, Italy 3. IBFM-CNR, University of Milano-Bicocca, Scientific Institute H San Raffaele, Segrate (MI), Italy 4. Department of Material Sciences, University of Milano-Bicocca, Milan, Italy 5. Department of Organic and Industrial Chemistry, University of Milano, Milan, Italy 6. Nerviano Medical Science srl, Nerviano (MI) Italy (literal)

Titolo

Choline and phosphatidylcholine fluorescent derivatives localization in carcinoma cells studied by laser scanning confocal fluorescence microscopy. (literal)

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. (literal)

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