Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells
Hexavalent chromium (Cr(VI)) is a serious environmental pollutant and human toxicant. Mammalian cells are very sensitive to chromate as they lack efficient chromate detoxifying strategy, e.g., chromate-reducing genes that are widely present in prokaryotes. To test whether introduction of prokaryotic...
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doaj-4e65235de53b40c5958985d2e67344a22020-11-24T21:19:11ZengMDPI AGInternational Journal of Molecular Sciences1422-00672015-05-01166118921190210.3390/ijms160611892ijms160611892Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 CellsXuan Liu0Gaofeng Wu1Yanli Zhang2Dan Wu3Xiangkai Li4Pu Liu5Department of Developmental Biology, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaDepartment of Developmental Biology, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaDepartment of Developmental Biology, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaDepartment of Developmental Biology, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaKey Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaDepartment of Developmental Biology, School of Life Sciences, Lanzhou University, Lanzhou 730000, ChinaHexavalent chromium (Cr(VI)) is a serious environmental pollutant and human toxicant. Mammalian cells are very sensitive to chromate as they lack efficient chromate detoxifying strategy, e.g., chromate-reducing genes that are widely present in prokaryotes. To test whether introduction of prokaryotic chromate-reducing gene into mammalian cells could render higher chromate resistance, an Escherichia coli chromate-reducing gene yieF was transfected into human HepG2 cells. The expression of yieF was measured in stably transfected cells HepG2-YieF by quantitative RT-PCR and found up-regulated by 3.89-fold upon Cr(VI) induction. In chromate-reducing ability test, HepG2-YieF cells that harbored the reductase showed significantly higher reducing ability of Cr(VI) than HepG2 control cells. This result was further supported by the evidence of increased Cr(VI)-removing ability of crude cell extract of HepG2-YieF. Moreover, HepG2-YieF demonstrated 10% higher viability and decreased expression of GSH synthesizing enzymes under Cr(VI) stress. Subcellular localization of YieF was determined by tracing GFP-YieF fusion protein that was detected in both nucleus and cytoplasm by laser confocal microscopy. Altogether, this study successfully demonstrated that the expression of a prokaryotic Cr(VI)-reducing gene yieF endowed mammalian cell HepG2 with enhanced chromate resistance, which brought new insight of Cr(VI) detoxification in mammalian cells.http://www.mdpi.com/1422-0067/16/6/11892chromate reductionyieFHepG2transfectionCr(VI) resistance |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Xuan Liu Gaofeng Wu Yanli Zhang Dan Wu Xiangkai Li Pu Liu |
spellingShingle |
Xuan Liu Gaofeng Wu Yanli Zhang Dan Wu Xiangkai Li Pu Liu Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells International Journal of Molecular Sciences chromate reduction yieF HepG2 transfection Cr(VI) resistance |
author_facet |
Xuan Liu Gaofeng Wu Yanli Zhang Dan Wu Xiangkai Li Pu Liu |
author_sort |
Xuan Liu |
title |
Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells |
title_short |
Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells |
title_full |
Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells |
title_fullStr |
Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells |
title_full_unstemmed |
Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells |
title_sort |
chromate reductase yief from escherichia coli enhances hexavalent chromium resistance of human hepg2 cells |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2015-05-01 |
description |
Hexavalent chromium (Cr(VI)) is a serious environmental pollutant and human toxicant. Mammalian cells are very sensitive to chromate as they lack efficient chromate detoxifying strategy, e.g., chromate-reducing genes that are widely present in prokaryotes. To test whether introduction of prokaryotic chromate-reducing gene into mammalian cells could render higher chromate resistance, an Escherichia coli chromate-reducing gene yieF was transfected into human HepG2 cells. The expression of yieF was measured in stably transfected cells HepG2-YieF by quantitative RT-PCR and found up-regulated by 3.89-fold upon Cr(VI) induction. In chromate-reducing ability test, HepG2-YieF cells that harbored the reductase showed significantly higher reducing ability of Cr(VI) than HepG2 control cells. This result was further supported by the evidence of increased Cr(VI)-removing ability of crude cell extract of HepG2-YieF. Moreover, HepG2-YieF demonstrated 10% higher viability and decreased expression of GSH synthesizing enzymes under Cr(VI) stress. Subcellular localization of YieF was determined by tracing GFP-YieF fusion protein that was detected in both nucleus and cytoplasm by laser confocal microscopy. Altogether, this study successfully demonstrated that the expression of a prokaryotic Cr(VI)-reducing gene yieF endowed mammalian cell HepG2 with enhanced chromate resistance, which brought new insight of Cr(VI) detoxification in mammalian cells. |
topic |
chromate reduction yieF HepG2 transfection Cr(VI) resistance |
url |
http://www.mdpi.com/1422-0067/16/6/11892 |
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