کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1910848 | 1046789 | 2008 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Novel enrichment of tumor cell transfectants expressing high levels of type 4 glutathione peroxidase using 7α-hydroperoxycholesterol as a selection agent
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کلمات کلیدی
7α-OOH3-(4,5-dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromideBHT7α-OHDFOGPx4GPx1DesferrioxaminedMPCDCPPBS1,2-dimyristoyl-sn-glycero-3-phosphocholine - 1،2-dimyristoyl-sn-glycero-3-phosphosocholineMTT - MTTt-BuOOH - T-BuOOHAntioxidant enzyme - آنزیم آنتی اکسیدانTransfection - ترانسفکشنdicetylphosphate - دیسیتیل فسفاتTumor cells - سلول های تومورtert-butyl hydroperoxide - هیدروپراکسید تری بوتیلCholesterol hydroperoxide - هیدروپراکسید کلسترولbutylated hydroxytoluene - هیدروکسی تورولین باتلاقیcholesterol - کلسترولglutathione reductase - گلوتاتیون ردوکتاز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
سالمندی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
A novel approach for selecting high expressing cells out of a general population that had been transfected with a construct encoding cytosolic type 4 glutathione peroxidase (GPx4) is reported. The approach is described for GPx4-null COH-BR1 breast tumor cells and is based on use of a highly specific GPx4 substrate, 7α-hydroperoxycholesterol (7α-OOH), as a selection agent. Cells recovering from a highly toxic dose of liposomal 7α-OOH were found to be substantially more resistant to a second 7α-OOH challenge than cells recovering from a less toxic dose, but were much less resistant to t-butyl hydroperoxide (t-BuOOH) or H2O2. Several clones isolated from the general transfectant population exhibited variable, relatively low GPx4 activities. However, clones from the 7α-OOH-selected population exhibited uniformly high GPx4 activities (each â¼3-fold higher than that of the starting transfectant population) and elevated steady-state mRNA levels. t-BuOOH could also be used for selecting high GPx4-expressing cells, but consistent recovery from toxic doses was more difficult than with 7α-OOH. Compared with conventional “hit or miss” cloning procedures, the 7α-OOH approach we describe affords a uniform population of high GPx4-activity cells in a relatively rapid manner. This approach should prove valuable for investigators interested in the peroxide regulatory properties of GPx4, in the context of both cytoprotection and redox signaling.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Free Radical Biology and Medicine - Volume 45, Issue 5, 1 September 2008, Pages 700-707
Journal: Free Radical Biology and Medicine - Volume 45, Issue 5, 1 September 2008, Pages 700-707
نویسندگان
Tamas Kriska, Vladislav V. Levchenko, Fong-Fong Chu, R. Steven Esworthy, Albert W. Girotti,