Correlating NAD(P)H lifetime shifts to tamoxifen resistance in breast cancer cells:A metabolic screening study with time-resolved flow cytometry  

作　　者：Samantha Valentino Karla Ortega-Sandoval Kevin D.Houston Jessica P.Houston 

机构地区：[1]Chemical and Materials Engineering,New Mexico State University 1040 S Horseshoe Dr.,Las Cruces,NM 88003,USA [2]Chemistry and Biochemistry,New Mexico State University 1175 N Horseshoe Dr.,Las Cruces,NM 88003,USA

出　　处：《Journal of Innovative Optical Health Sciences》2025年第1期101-115,共15页创新光学健康科学杂志(英文)

基　　金：the National Institute of Health for supporting this research under grants NIH R35GM152076,NIH 1SC1GM127175-01,NIH T32GM148394.

摘　　要：Time-resolved flow cytometry(TRFC)was used to measure metabolic differences in estrogen receptor-positive breast cancer cells.This specialty cytometry technique measures fluorescence lifetimes as a single-cell parameter thereby providing a unique approach for high-throughput cell counting and screening.Differences in fluorescence lifetime were detected and this was associated with sensitivity to the commonly prescribed therapeutic tamoxifen.Differences in fluorescence lifetime are attributed to the binding states of the autofluorescent metabolite NAD(P)H.The function of NAD(P)H is well described and in general involves cycling from a reduced to oxidized state to facilitate electron transport for the conversion of pyruvate to lactate.NAD(P)H fluorescence lifetimes depend on the bound or unbound state of the metabolite,which also relates to metabolic transitions between oxidative phosphorylation and glycolysis.To determine if fundamental metabolic profiles differ for cells that are sensitive to tamoxifen compared to those that are resistant,large populations of MCF-7 breast cancer cells were screened and fluorescence lifetimes were quantified.Additionally,metabolic differences associated with tamoxifen sensitivity were measured with a Seahorse HS mini metabolic analyzer(Agilent Technologies Inc.Santa Clara,CA)and confocal imaging.Results show that tamoxifen-resistant breast cancer cells have increased utilization of glycolysis for energy production compared to tamoxifen-sensitive breast cancer cells.This work is impacting because it establishes an early step toward developing a reliable screening technology in which large cell censuses can be differentiated for drug sensitivity in a label-free fashion.

关 键 词：TIME-RESOLVED flow cytometry AUTOFLUORESCENCE fluorescence lifetime breast cancer metabolism 

分 类 号：R737.9[医药卫生—肿瘤]