Caco-2细胞中药物转运蛋白的mRNA表达及活力评价

目的 系统研究Caco-2细胞中各药物转运蛋白的mRNA表达水平及转运活力，对比其与人正常肠道中药物转运蛋白表达的差异。方法 实时荧光定量PCR（qRT-PCR）方法测定Caco-2细胞中人肠道相关转运蛋白MDR1、BCRP、MRP2、OATP1A2、OATP2B1和PEPT1的表达水平；将Caco-2细胞接种于Transwell板内培养21 d并给予不同药物转运蛋白的底物及抑制剂，评价Caco-2细胞中相关转运蛋白的转运活性。结果 qRT-PCR结果表明，药物转运蛋白MDR1、MRP2、BCRP和OATP2B1在Caco-2细胞中均有相对高的表达，表达量的顺序为：MDR1 > MRP2 > OATP2B1 > BCRP，在正常人肠道表达量顺序为BCRP > MDR1 > MRP2 > OATP2B1；转运蛋白活力评价表明，各药物转运蛋白的活力测试结果均为阳性，验证了基因的表达水平结果。结论 Caco-2细胞中表达正常人体肠道表达的部分药物转运蛋白（MDR1、MRP2、BCRP和OATP2B1），表达水平与正常人体肠道中大致相当，但也存在一定差异。

Expression of drug transporters mRNA in Caco-2 cells and its viability evaluation

Objective To study the mRNA expression and transport activity of drug transporters in Caco-2 cells, and compare the differences of the expression of drug transporters between Caco-2 cells and human normal intestinal. Method Establishing a method for the determination of mRNA expression of drug transporter by qRT-PCR method to determine the expression levels of human intestinal related transport proteins MDR1, BCRP, MRP2, OATP1A2, OATP2B1, and PEPT1 in Caco-2 cells; Caco-2 was then seeded in Transwell plates for 21 days and administered with different drug transporter substrates and inhibitors to assess the transporter activity of the relevant transporter in Caco-2 cells. Results The qRT-PCR results indicated that MDR1, MRP2, BCRP, and OATP2B1 were relatively higher expressed in Caco-2 cells. The relative expression levels of various drug transporters were as follows:MDR1 > MRP2 > OATP2B1 > BCRP, the expression level of intestinal tract in normal subjects was BCRP > MDR1 > MRP2 > OATP2B1. Drug transporter activity assessment showed that all drug transporter activity test results were positive, validating the expression level of the previous drug transporter gene. Conclusions The expression levels of many drug transporters (MDR1, MRP2, BCRP and OATP2B1) originated from normal human intestinal tract expressed in Caco-2 cells were roughly equivalent to those expressed in the normal human intestinal tract, but there were also some differences between them.