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Effect of histone Deacetylase Inhibition on the Expression of multidrug resistance-associated protein 2 in a Human Placental Trophoblast Cell Line

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Chinese Medical Journal 2017年第11期130卷 1352-1360页

作者： Hong-Yu Duan Dan Ma Kai-Yu Zhou Tao Wang Yi Zhang Yi-Fei Li Jin-Lin Wu Yi-Min Hua Chuan Wang Department of Pediatric Cardiology West China Second University Hospital Sichuan University Chengdu Sichuan 610041 China The Cardiac Development and Early Intervention Unit West China Institute of Women and Children's Health West China Second University Hospital Sichuan University Chengdu Sichuan 610041 China Department of Pediatric Rehabilitation West China Second University Hospital Sichuan University Chengdu Sichuan 610041 China Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University) Ministry of Education Chengdu Sichuan 610041 China Key Laboratory of Development and Diseases of Women and Children of Sichuan Province West China Second University Hospital Sichuan University Chengdu Sichuan 610041 China Department of Pediatrics West China Second University Hospital Sichuan University Chengdu Sichuan 610041 China

Background： Placental multidrug resistance-associated protein 2 （MRP2）, encoded by ABCC2 gene in human, plays a significant role in regulating drugs＇ transplacental transfer rates. Studies o11 placental MRP2 regulation could provide more therapeutic targets for individualized and safe pharmacotherapy during pregnancy. Currently, the roles of epigenetic mechanisms in regulating placental drug transporters are still unclear. This study aimed to investigate the effect of histone deacetylases （HDACs） inhibition on MRP2 expression in the placental trophoblast cell line and to explore whether HDAC 1/2/3 are preliminarily involved in this process. Methods： The human choriocarcinoma-derived trophoblast cell line （Bewo cells） was treated with the HDAC inhibitors-trichostatin A （TSA） at different concentration gradients of 0.5, 1.0, 3.0, and 5.0 μmol/L. Cells were harvested after 24 and 48 h treatment. Small interfering RNA （siRNA） specific for HDACI/HDAC2/HDAC3 or control siRNA was transfected into cells. Total HDAC activity was detected by colorimetric assay kits. HDAC 1/2/3/ABCC2 messenger RNA （mRNA） and protein expressions were determined by real-time quantitative polymerase chain reaction and Western-blot analysis, respectively. Immunofluorescence for MRP2 protein expression was visualized and assessed using an immunofluorescence microscopy and ImageJ software, respectively. Results： TSA could inhibit total HDAC activity and HDAC 1/2/3 expression in company with increase ofM RP2 expression in Bewo cells. Reduction of HDAC 1 protein level was noted after 24 h of TSA incubation at 1.0, 3.0, and 5.0 μmol/L （vs. vehicle group, all P 〈 0.001 ）, accompanied with dose-dependent induction of MRP2 expression （P = 0.045 for 1.0 μmol/L, P = 0.001 for 3.0 μmol/L, and P 〈 0.001 for 5.0 μmol/L）, whereas no significant diferences in MRP2 expression were noted after HDAC2/3 silencing. Fluorescent micrograph images of MRP2 protein were expressed on the cell membrane. The fluorescent inte

关键词： epigenetic regulation: histone deacetylases: multidrug resistance-associated protein 2 Placenta

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