Dicoumarol enhances gemcitabine-induced cytotoxicity in high NQ01-expressing cholangiocarcinoma cells
Dicoumarol enhances gemcitabine-induced cytotoxicity in high NQ01-expressing cholangiocarcinoma cells
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DOI：
10.3748/wjg.v16.i19.2362
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发表时间：
2010-05-21
影响因子：
4.3
通讯作者：
Kukongviriyapan, Veerapol
中科院分区：
文献类型：
--
作者：
Buranrat, Benjaporn;Prawan, Auemduan;Kukongviriyapan, Veerapol
目的：为了研究NAD（P）H醌氧化还原酶-1（NQO 1）的有效抑制剂双香豆素（dicoumarol）是否能增强吉西他滨对胆管癌细胞（CCA）的细胞毒性，以及活性氧的产生在致敏CCA中的作用。方法：使用4种具有不同NQ 01活性的人CCA细胞系：KKU-100、KKU-OCA 17、KKU-M214和Chang肝细胞。测定NQ 01活性和mRNA表达。在用吉西他滨处理之前，用相关浓度的双香豆素预处理细胞。通过荧光染料染色测定细胞毒性。通过测定细胞谷胱甘肽水平和使用二氢荧光素二乙酸酯产生活性氧来检查氧化剂形成。用JC-1荧光探针测定线粒体跨膜电位。结果：双香豆素可显著增强吉西他滨对高NQ 01活性和mRNA表达细胞KKU-100和KKU-OCA 17的细胞毒作用，但对低NQ 01活性细胞的细胞毒作用不明显。与KKU-M214细胞相比，双香豆素可诱导KKU-100细胞中谷胱甘肽的细胞氧化还原显着降低。在抑制NQ 01活性的浓度下，双香豆素并不改变线粒体跨膜电位和活性氧的产生。吉西他滨单独诱导NF-κ B和Bcl-xL蛋白表达的活化。结论：NQ 01可能是CCA的重要增敏因子，抑制NQ 01可能是治疗CCA的一种策略。(C)2010年百世登。All rights reserved.
AIM: To investigate whether dicoumarol, a potent inhibitor of NAD(P)H quinone oxidoreductase-1 (NQO1), potentiates gemcitabine to induce cytotoxicity in cholangiocarcinoma cells (CCA) and the role of reactive oxygen generation in sensitizing the cells.METHODS: Four human cell lines with different NQ01 activity were used; the human CCA cell lines, KKU-100, KKU-OCA17, KKU-M214, and Chang liver cells. NQ01 activity and mRNA expression were determined. The cells were pretreated with dicoumarol at relevant concentrations before treatment with gemcitabine. Cytotoxicity was determined by staining with fluorescent dyes. Oxidant formation was examined by assay of cellular glutathione levels and reactive oxygen species production by using dihydrofluorescein diacetate. Measurement of mitochondrial transmembrane potential was performed by using JC-1 fluorescent probe. Western blotting analysis was performed to determine levels of survival related proteins.RESULTS: Dicoumarol markedly enhanced the cytotoxicity of gemcitabine in KKU-100 and KKU-OCA17, the high NQ01 activity and mRNA expressing cells, but not in the other cells with low NQ01 activity. Dicoumarol induced a marked decrease in cellular redox of glutathione in KKU-100 cells, in contrast to KKU-M214 cells. Dicoumarol at concentrations that inhibited NQ01 activity did not alter mitochondrial transmembrane potential and production of reactive oxygen species. Gemcitabine alone induced activation of NF-kappa B and Bcl-xL protein expression. However, gemcitabine and dicoumarol combination induced increased p53 and decreased Bcl-xL levels in KKU-100, but not in KKU-M214 cells.CONCLUSION: NQ01 may be important in sensitizing cells to anticancer drugs and inhibition of NQ01 may be a strategy for the treatment of CCA. (C) 2010 Baishideng. All rights reserved.