Duan Hongyu, Wang Chuan, Zhou Kaiyu, Wang Tao, Li Yifei, Qiu Dajian, Li Qiang, Zhang Yi, Hua Yimin
Department of Pediatric Cardiology, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, 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, China; West China Medical School of Sichuan University, Chengdu, Sichuan, China.
Department of Pediatric Cardiology, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, 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, China.
Placenta. 2017 Jan;49:37-47. doi: 10.1016/j.placenta.2016.11.011. Epub 2016 Nov 23.
Placental P-glycoprotein (P-gp), encoded by ABCB1 gene in human, plays a significant role in regulating drugs' transplacental transfer rates. Investigations on placental P-gp regulation could provide more therapeutic targets for individualized and safe pharmacotherapy during pregnancy. Currently, the epigenetic control of placental P-gp is rare. This study aimed to investigate the effect of histone deacetylases (HDACs) inhibition on P-gp expression in placental trophoblast cell lines and to explore whether HDAC1/2/3 was involved in this process preliminarily.
Human placental trophoblast cell lines (Bewo and JAR) were treated with two different HDAC inhibitors-suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA) at different concentration gradients of 0.5, 1.0, 3.0 and 5.0 μM. Cells were harvested after 24, 48, and 72 h treatment. Total HDAC activity was detected by colorimetric assay Kits. HDAC1/2/3/ABCB1 mRNA and protein expressions were determined by real-time quantitative PCR and western-blot, respectively. Pearson correlation analysis test was performed to explore the relationship between HDAC1/2/3 mRNA and ABCB1 mRNA expression.
SAHA and TSA could inhibit total HDAC activity and placental HDAC1/2/3 expression both in Bewo and JAR, but displayed a transient induction of HDAC mRNA or protein level after being treated at low dosage or prolonged exposure to drugs. Discordance in HDAC mRNA and protein expression was also observed. Placental P-gp expression was significantly induced in company with HDACs inhibition. There was a significant negative linear relationship between HDAC1/2 mRNA and ABCB1 mRNA expression.
HDACs inhibition could up-regulate placental P-gp expression in trophoblast cells, and HDAC1/2 was most likely to be involved in this process.
人ABCB1基因编码的胎盘P-糖蛋白（P-gp）在调节药物经胎盘转运速率方面发挥着重要作用。对胎盘P-gp调控的研究可为孕期个体化和安全药物治疗提供更多治疗靶点。目前，关于胎盘P-gp的表观遗传控制研究较少。本研究旨在探讨组蛋白去乙酰化酶（HDACs）抑制对胎盘滋养层细胞系中P-gp表达的影响，并初步探究HDAC1/2/3是否参与此过程。
用人胎盘滋养层细胞系（Bewo和JAR）分别用两种不同的HDAC抑制剂——异羟肟酸苯丁酯（SAHA）和曲古抑菌素A（TSA），浓度梯度为0.5、1.0、3.0和5.0 μM进行处理。处理24、48和72小时后收获细胞。用比色法试剂盒检测总HDAC活性。分别用实时定量PCR和蛋白质免疫印迹法测定HDAC1/2/3/ABCB1 mRNA和蛋白表达。进行Pearson相关分析以探究HDAC1/2/3 mRNA与ABCB1 mRNA表达之间的关系。
SAHA和TSA均可抑制Bewo和JAR中的总HDAC活性及胎盘HDAC1/2/3表达，但在低剂量处理或长时间暴露于药物后会短暂诱导HDAC mRNA或蛋白水平。还观察到HDAC mRNA和蛋白表达不一致。伴随HDACs抑制，胎盘P-gp表达显著上调。HDAC1/2 mRNA与ABCB1 mRNA表达之间存在显著的负线性关系。
HDACs抑制可上调滋养层细胞中胎盘P-gp表达，且HDAC1/2最有可能参与此过程。
