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001-es BibID:	BIBFORM004876
035-os BibID:	WOS:000228153700002
Első szerző:	Megyeri Attila (orvos)
Cím:	Development of a stereological method to measure levels of fluoropyrimidine metabolizing enzymes in tumor sections using laser scanning cytometry / Attila Megyeri, Zsolt Bacsó, Anthony Shields, James F. Eliason
Dátum:	2005
ISSN:	1552-4922
Megjegyzések:	The enzymes thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) influence the activities of fluoropyrimidine anticancer drugs. The sensitivity of cancer cells to capecitabine, which is an oral, tumor-selective pre-prodrug of 5-fluorouracil may correlate better to the TP/DPD ratio than to levels of either enzyme alone. Our goal was to develop a quantitative immunofluorescent method for estimating the levels of TP, DPD, and their ratio in archival tumor sections. METHODS: Mouse anti-TP and rat anti-DPD monoclonal antibodies were used for parallel indirect immunofluorescent staining. The fluorescence was measured using a laser scanning cytometer (LSC; CompuCyte, Cambridge, MA) in single cells and in sections prepared from cell lines and a human tumor. The phantom contouring feature of the LSC provided a stereologic approach for collecting the fluorescence intensity data from sections. RESULTS: The relative fluorescence intensities measured in single cells or in sections of the cell lines, using single or double labeling, were similar, supporting the suitability of phantom contouring and two-color staining. Sections of the T-24 and ZR-75-1 cell lines placed on the same slide as the tumor section were used as internal standards for fluorescence measurements. The TP/DPD ratios measured in three cell lines correlated well with the cytotoxicity of 5'-deoxy-5-fluorouridine measured in vitro, indicating that the measurements are related to the biological activity of the drug. CONCLUSIONS: Plotting the data as contour maps of the topologic distribution of fluorescence intensities in tumor sections allows subsequent histopathologic examination, which may reveal features of the tumors leading to high or low ratios of these enzymes. In addition, this method can be used for any drug target/metabolic system where the key components are known and suitable antibodies are available
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban analysis Antibodies Antibodies,Monoclonal Antimetabolites,Antineoplastic Cell Line Cell Line,Tumor Cell Proliferation Cells chemistry Dihydrouracil Dehydrogenase (NADP) drug effects Dyes Enzymes enzymology Esophageal Neoplasms Floxuridine Fluorescence Fluorescent Antibody Technique Fluorescent Dyes Human Humans In Vitro Laser Scanning Cytometry metabolism methods Neoplasms pathology pharmacology Pyrimidines Research Support Thymidine Thymidine Phosphorylase egyetemen (Magyarországon) készült közlemény
Megjelenés:	Cytometry. Part A. - 64 : 2 (2005), p. 62-71. -
További szerzők:	Bacsó Zsolt (1963-) (biofizikus) Shields, Anthony Eliason, James F.
Internet cím:	elektronikus változat DOI
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