%0 Journal Article
%T Enzyme-mediated depletion of l-cyst(e)ine synergizes with thioredoxin reductase inhibition for suppression of pancreatic tumor growth
%A Achinto Saha
%A Everett Stone
%A George Georgiou
%A John DiGiovanni
%A Paul Gries
%A Sabin Kshattry
%A Stefano Tiziani
%J Archive of "NPJ Precision Oncology".
%D 2019
%R 10.1038/s41698-019-0088-z
%X Effect of cyst(e)inase treatment on survival of PDAC cell lines. a Relative cell survival 48ˋh after cyst(e)inase treatment in three pancreatic cancer cell lines (nˋ=ˋ3 cultures for each dose). b Intracellular l-Cys levels upon treatment with 250ˋnM cyst(e)inase (one treated and one untreated culture for each time point, data from a representative experiment). c Intracellular GSH levels 24ˋh after treatment (nˋ=ˋ3 cultures for each dose). d Cellular ROS levels as assessed by 2∩,7∩-dichlorofluorescin diacetate (DCFDA) fluorescence 24ˋh after treatment. tert-Butyl hydrogen peroxide (TBHP) treatment is included as a positive control for ROS accumulation (nˋ=ˋ3每5 independent experiments). e Mitochondrial ROS levels as assessed by MitoSOX fluorescence following 24ˋh cyst(e)inase treatment (nˋ=ˋ3每6 independent experiments). f Oxidative stress signaling via JNK and ATM, and apoptosis signaling following cyst(e)inase treatment. Cys cyst(e)inase. A white line is added to separate the TBHP treatment group with the rest of the time course data. Original blots can be found in the Supplementary Figures. g Cell death in Panc1 cells as assessed by trypan blue staining 48ˋh after treatment (nˋ=ˋ3 independent experiments). h Cell death in MIA-PaCa2 and BxPC3 cells 72ˋh after cyst(e)inase treatment (nˋ=ˋ3每4 independent experiments). i Cellular growth assay of Panc1, MIA-PaCa2, and BxPC3 cells (nˋ=ˋ3 cultures for each time point) with cyst(e)inase treatment. For f, ※+§ represents 250ˋnM cyst(e)inase treatment. All data represent meanˋ㊣ˋs.e.m. except for b. Dotted line in a represents the 95% confidence band for the fitted curve (four-parameter logistic model). *Pˋ<ˋ0.05, ****Pˋ<ˋ0.0001; compared to untreated controls (c每e, g, h), or between the bottom plateaus of the fitted curve (a); two-way ANOVA (c每e) or one-way ANOVA (a, g, h) with Bonferroni＊s method for multiple-comparison test. Experiments in f were repeated twice with similar results. All other experiments were repeated three times or mor
%K Pancreatic cancer
%K Biologics
%K Cancer therapy
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6546752/