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Int. J. Mol. Sci. 2015, 16(6), 11892-11902; doi:10.3390/ijms160611892

Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells

1,†
,
1,†
,
1
,
1
,
2
and
1,*
1
Department of Developmental Biology, School of Life Sciences, Lanzhou University, Lanzhou 730000, China
2
Key Laboratory of Cell Activities and Stress Adaptations, School of Life Sciences, Lanzhou University, Lanzhou 730000, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Ji-Dong Gu
Received: 9 February 2015 / Revised: 6 May 2015 / Accepted: 13 May 2015 / Published: 26 May 2015
View Full-Text   |   Download PDF [1749 KB, uploaded 26 May 2015]   |  

Abstract

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. View Full-Text
Keywords: chromate reduction; yieF; HepG2; transfection; Cr(VI) resistance chromate reduction; yieF; HepG2; transfection; Cr(VI) resistance
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Liu, X.; Wu, G.; Zhang, Y.; Wu, D.; Li, X.; Liu, P. Chromate Reductase YieF from Escherichia coli Enhances Hexavalent Chromium Resistance of Human HepG2 Cells. Int. J. Mol. Sci. 2015, 16, 11892-11902.

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