Effect of chlorpyrifos on efflux transporter gene expression and function in Caco-2 cells
Effect of chlorpyrifos on efflux transporter gene expression and function in Caco-2 cells
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DOI：
10.1016/j.tiv.2003.12.006
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发表时间：
2004-08-01
影响因子：
3.2
通讯作者：
Cook, TJ
中科院分区：
文献类型：
--
作者：
Agarwala, S;Chen, W;Cook, TJ
研究了毒死蜱（CPF）及其代谢产物毒死蜱氧磷（CPO）对Caco-2细胞多药耐药1（MDR 1）基因表达和外排转运蛋白功能的影响。使用RT-PCR和竞争性PCR（compPCR）技术测试CPF和CPO对Caco-2细胞中基因表达的影响作为时间的函数。RT-PCR结果显示CPF暴露对MDR 1表达的影响在8h达到最大，24 h后降低。用CPO的研究显示仅在4小时表达初始增加。用CPF处理组进行compPCR测定以定量基因表达水平的变化。compPCR数据证实并定量了使用半定量RT-PCR的时程研究的结果。除了基因表达研究外，还使用在Transwell（TM）板中的半透膜上生长的Caco-2细胞研究了外排转运蛋白功能的变化。在用CPF处理8 h的细胞中测定维拉帕米的渗透性，外排率表明，与对照组相比，维拉帕米以更高的速率从CPF处理的细胞中外排，证实了CPF对转运蛋白功能的诱导作用。这些结果表明，CPF有可能通过改变膜外排转运蛋白的表达和功能来调节药物的生物利用度（C）2003 Elsevier Ltd.保留所有权利。
The effect of chlorpyrifos (CPF) and its metabolite, chlorpyrifos-oxon (CPO), on multidrug resistance-1 (MDR 1) gene expression and efflux transporter function in Caco-2 cells was determined. The effect of CPF and CPO on gene expression in Caco-2 cells was tested as a function of time using RT-PCR and competitive PCR (compPCR) techniques. The RT-PCR results depicted a maximal effect of CPF exposure on MDR1 expression at 8 h, which decreased at 24 h. Studies with CPO displayed an initial increase in expression at 4 h only. The compPCR assays were conducted with the CPF-treated group to quantify the changes in gene expression levels. The compPCR data confirmed and quantitated the results from the time-course study using semiquantitative RT-PCR. In addition to the gene expression studies, changes in efflux transporter function were investigated using Caco-2 cells grown on semipermeable membranes in Transwell(TM) plates. The permeability of verapamil was determined in cells treated for 8 h with CPF, Efflux ratios demonstrated that verapamil was effluxed at a higher rate from the CPF-treated cells as compared to the control group, confirming the inductive action of CPF on transporter function. These results suggest that CPF has the potential to modulate the bioavailability of drugs via changes in expression and function of membrane efflux transporters' (C) 2003 Elsevier Ltd. All rights reserved.