Cabozantinib reverses multidrug resistance of human hepatoma HepG2/adr cells by modulating the function of P-glycoprotein
Cabozantinib reverses multidrug resistance of human hepatoma HepG2/adr cells by modulating the function of P-glycoprotein
复制标题
卡博替尼通过调节 P-糖蛋白的功能逆转人肝癌 HepG2/adr 细胞的多药耐药性。
DOI：
10.1111/liv.12524
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发表时间：
2015-03-01
影响因子：
6.7
通讯作者：
Shang, Chang-zhen
中科院分区：
文献类型：
--
作者：
Xiang, Qing-feng;Zhang, Dong-mei;Shang, Chang-zhen
背景与目的卡博替尼是一种小分子多靶点酪氨酸激酶抑制剂，已进入治疗肝细胞癌（HCC）的III期临床试验。本研究评估了卡博替尼逆转P-糖蛋白（P-gp）介导的多药耐药（MDR）的机制。方法采用CCK-8法和肿瘤移植物分别在体外和体内研究了卡博替尼逆转MDR的作用。通过荧光显微镜和流式细胞术评价底物保留测定。Western blotting检测蛋白表达水平。通过qPCR测定mRNA表达。使用Pgp-Glo测定系统研究P-gp的ATP酶活性。卡博替尼与P-gp在分子水平上的结合机制进行了评估，使用对接analysis.ResultsCabozantinib增强HepG 2/adr和HEK 293-MDR 1细胞中P-gp底物药物的细胞毒性，但对非P-gp底物没有影响。此外，卡博替尼增加了HepG 2/adr细胞中P-gp底物的积累，但对HepG 2细胞没有影响。此外，卡博替尼不改变P-gp mRNA或蛋白质的表达，但刺激P-gp ATP酶的活性。对接研究表明，卡博替尼和维拉帕米可能部分共享P-gp上的结合位点。卡博替尼的逆转浓度不影响MET、AKT和ERK 1/2的表达。显著地，cabozantinib增加阿霉素在P-gp过表达的HepG 2/adr细胞裸鼠移植瘤中的抑制功效。结论cabozantinib通过直接抑制P-gp的外排功能逆转P-gp介导的MDR，表明cabozantinib可能有助于逆转P-gp介导的MDR在HCC和其他癌症化疗中。
Background & AimsCabozantinib, a small-molecule multitargeted tyrosine kinase inhibitor, has entered into a phase III clinical trial for the treatment of hepatocellular carcinoma (HCC). This study assessed the mechanistic effect of cabozantinib on the reversal of P-glycoprotein (P-gp)-mediated multidrug resistance (MDR).MethodsCCK-8 assays and tumour xenografts were used to investigate the reversal of MDR in vitro and in vivo respectively. Substrate retention assays were evaluated by fluorescence microscope and flow cytometry. Western blotting was used to detect protein expression levels. mRNA expression was determined by qPCR. The ATPase activity of P-gp was investigated using Pgp-Glo assay systems. The binding mechanism of cabozantinib to P-gp at the molecular level was evaluated using docking analysis.ResultsCabozantinib enhanced the cytotoxicity of P-gp substrate drugs in HepG2/adr and HEK293-MDR1 cells but had no effect on non-P-gp substrates. In addition, cabozantinib increased the accumulation of P-gp substrates in HepG2/adr cells but had no effect in HepG2 cells. Furthermore, cabozantinib did not alter the expression of P-gp mRNA or protein but did stimulate the activity of P-gp ATPase. The docking study indicated that cabozantinib and verapamil may partially share a binding site on P-gp. The reversal concentrations of cabozantinib did not affect the expression of MET, AKT and ERK1/2. Significantly, cabozantinib increased the inhibitory efficacy of doxorubicin in P-gp-overexpressing HepG2/adr cell xenografts in nude mice.ConclusionCabozantinib reverses P-gp-mediated MDR by directly inhibiting the efflux function of P-gp, indicating that cabozantinib may help to reverse P-gp-mediated MDR in HCC and other cancer chemotherapy.