Prokaryotic Arsenate Reductase Enhances Arsenate Resistance in Mammalian Cells

Title:Prokaryotic Arsenate Reductase Enhances Arsenate Resistance in Mammalian Cells

Volume: 6 Issue: 2

Author(s): Dan Wu, Xuanyu Tao, Gaofeng Wu, Xiangkai Li and Pu Liu

Affiliation:

Keywords: Arsenate reductase, arsenate resistance, detoxification, HepG₂, transfection.

Abstract: Arsenic is a well-known heavy metal toxicant in the environment. Bioremediation of heavy metals has been proposed as a low-cost and eco-friendly method. This article described some of recent patents on transgenic plants with enhanced heavy metal resistance. Further, to test whether genetic modification of mammalian cells could render higher arsenic resistance, a prokaryotic arsenic reductase gene arsC was transfected into human liver cancer cell HepG₂. In the stably transfected cells, the expression level of arsC gene was determined by quantitative real-time PCR. Results showed that arsC was expressed in HepG₂ cells and the expression was upregulated by 3 folds upon arsenate induction. To further test whether arsC has function in HepG₂ cells, the viability of HepG₂-pCI-ArsC cells exposed to arsenite or arsenate was compared to that of HepG₂-pCI cells without arsC gene. The results indicated that arsC increased the viability of HepG₂ cells by 25% in arsenate, but not in arsenite. And the test of reducing ability of stably transfected cells revealed that the concentration of accumulated trivalent arsenic increased by 25% in HepG₂-pCI-ArsC cells. To determine the intracellular localization of ArsC, a fusion vector with fluorescent marker pEGFP-N1-ArsC was constructed and transfected into.HepG₂. Laser confocal microscopy showed that EGFP-ArsC fusion protein was distributed throughout the cells. Taken together, these results demonstrated that prokaryotic arsenic resistant gene arsC integrated successfully into HepG₂ genome and enhanced arsenate resistance of HepG₂, which brought new insights of arsenic detoxification in mammalian cells.

Cite this article as:

Wu Dan, Tao Xuanyu, Wu Gaofeng, Li Xiangkai and Liu Pu, Prokaryotic Arsenate Reductase Enhances Arsenate Resistance in Mammalian Cells, Recent Patents on Food, Nutrition & Agriculture 2014; 6 (2) . https://dx.doi.org/10.2174/2212798407666141210110248