Minor role of pregnane-x-receptor for acquired multidrug resistance in head and neck squamous cell carcinoma in vitro
Minor role of pregnane-x-receptor for acquired multidrug resistance in head and neck squamous cell carcinoma in vitro
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DOI：
10.1007/s00280-013-2133-x
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发表时间：
2013-05-01
影响因子：
3
通讯作者：
Theile, Dirk
中科院分区：
文献类型：
--
作者：
Pablo Rigalli, Juan;Reuter, Tasmin;Theile, Dirk
获得性多药耐药（MDR）与孕烷-X-受体（PXR）介导的药物代谢和转运蛋白的过度表达有关。为探讨PXR与头颈部鳞癌多药耐药的关系，本研究采用8株头颈部鳞癌细胞系，通过细胞增殖实验检测紫杉醇、顺铂和5-氟尿嘧啶（5-FU）对头颈部鳞癌的疗效，并通过实时荧光定量PCR、Western blotting和报告基因分析检测PXR的表达和活性。PXR敲低的方法，使用shRNA编码载体被施加到估计的作用PXR为天然MDR.Drug耐药性范围为5.2和620 nM之间的紫杉醇，4.5和58 μ M之间的顺铂，和1.1和5,467 μ M之间的5-FU。PXR mRNA表达的缺失主要伴随细胞色素P450 3A 4（CYP 3A 4）和P-糖蛋白（P-gp，ABCB 1）mRNA表达的缺失。PXR的mRNA和蛋白表达与耐药性无关。然而，PXR活性倾向于与紫杉醇的IC 50值相关（p = 0.08）。与乱序序列对照相比，一种细胞系中PXR的敲低对紫杉醇疗效有轻微但不显著的影响。令人惊讶的是，只有在两个细胞系中，PXR活性增加了众所周知的诱导剂rifampicin.This研究表明，PXR的故障，从而在HNSCC医源性化疗耐药的小的相关性。
Acquired multidrug resistance (MDR) has been linked to overexpression of drug-metabolising and transporting proteins mediated by pregnane-x-receptor (PXR). The aim of this work was to establish the relevance of PXR for MDR in head and neck squamous cell carcinoma (HNSCC).Using eight HNSCC cell lines, we determined the efficacy of paclitaxel, cisplatin and 5-fluorouracil (5-FU) via proliferation assays and determined the expression and activity of PXR through quantitative real-time polymerase chain reaction, western blotting and luciferase-based reporter gene assay. PXR knockdown approaches using shRNA-encoding vectors were applied to estimate the role of PXR for native MDR.Drug resistance ranged between 5.2 and 620 nM for paclitaxel, varied between 4.5 and 58 mu M for cisplatin, and varied between 1.1 and 5,467 mu M for 5-FU. Lack of PXR mRNA expression was mostly accompanied by the absence of mRNA expression of cytochrome P450 3A4 (CYP3A4) and P-glycoprotein (P-gp, ABCB1) expression. Neither mRNA nor protein expression of PXR correlated with drug resistance. However, PXR activity tended to correlate with IC50 values of paclitaxel (p = 0.08). Knockdown of PXR in one of the cell lines had a slight but not significant impact on paclitaxel efficacy compared to scrambled sequence control. Surprisingly, only in two cell lines, PXR activity was increased by the well-known inductor rifampicin.This study suggests a malfunctioning of PXR and thus a minor relevance for iatrogenic chemotherapy resistance in HNSCC.