Gene expression of CYP3A4, ABC-transporters (MDR1 and MRP1-MRP5) and hPXR in three different human colon carcinoma cell lines
Gene expression of CYP3A4, ABC-transporters (MDR1 and MRP1-MRP5) and hPXR in three different human colon carcinoma cell lines
复制标题
DOI：
10.1111/j.2042-7158.2003.tb02434.x
复制
发表时间：
2003-01-01
影响因子：
3.3
通讯作者：
Drewe, J
中科院分区：
文献类型：
--
作者：
Pfrunder, A;Gutmann, H;Drewe, J
结肠癌细胞系被广泛用作药物肠吸收的筛选模型。然而，重要的运输系统和代谢酶的表达尚未完全表征。研究了多药耐药基因1 （MDR1）和细胞色素P450异构体3A4 （CYP3A4）在Caco-2亲本、Caco-2 TC-7 （TC-7）和LS180细胞株中的表达和诱导作用。在相同的三个细胞系中，我们研究了多药耐药相关蛋白家族（MRP1-MRP5）和人妊娠X受体（hPXR）的表达，这可能对MDR1和CYP3A4的诱导很重要。用利福平或1,25（OH)(2)D-3）处理细胞72 h，提取总RNA。反转录实时聚合酶链反应（TaqMan）测定mRNA表达水平。我们已经证明，在LS180细胞中，两种诱导剂均可诱导MDR1和CYP3A4。在Caco-2亲本细胞和TC-7细胞中，CYP3A4仅能被1,25(OH)2D3诱导。此外，几种转运蛋白（MDR1、MRP1-MRP5）和CYP3A4在不同结肠癌来源细胞系中的基因表达也存在差异。hPXR mRNA在3种细胞系中均有表达，但在LS180细胞中检测到的mRNA量显著高于Caco-2和TC-7细胞。我们得出结论，LS180细胞是研究MDR1和CYP3A4诱导的合适模型，但对于药物转运研究，Caco-2亲代和TC-7细胞更适合作为更生理的模型。
Colon carcinoma cell lines are used widely as screening models for intestinal absorption of drugs. However, the expression of important transport systems and of metabolic enzymes is not completely characterized yet. The expression and inducibility of multidrug resistance gene 1 (MDR1) and cytochrome P450 isoform 3A4 (CYP3A4) was investigated in Caco-2 parental, Caco-2 TC-7 (TC-7) and LS180 cell lines. In the same three cell lines, we investigated the expression of isoforms of the multidrug resistance associated protein family (MRP1-MRP5) and the human pregnane X receptor (hPXR), which may be important for MDR1 and CYP3A4 induction. Cells were treated with rifampicin or 1alpha,25-dihydroxycholecalciferol (1,25(OH)(2)D-3) for 72 h and the total RNA was extracted. Afterwards reverse transcription real-time polymerase chain reaction (TaqMan) assay was performed to determine the mRNA expression level. We have shown that in LS180 cells, MDR1 and CYP3A4 were inducible with both inducers. In Caco-2 parental and TC-7 cells, CYP3A4 was only inducible with 1,25(OH)2D3. Furthermore, differences were shown in gene expression of several transport proteins (MDR1 and MRP1-MRP5) and CYP3A4 in different human colon carcinoma derived cell lines. hPXR mRNA was expressed in all three cell lines but the amount of mRNA detected was significantly higher in LS180 cells than in Caco-2 and TC-7 cells. We concluded that LS180 cells were a suitable model to study MDR1 and CYP3A4 induction, but for drug transport studies Caco-2 parental and TC-7 cells would be preferred as the more physiological model.