Can P-glycoprotein mediate resistance to nilotinib in human leukaemia cells?
Can P-glycoprotein mediate resistance to nilotinib in human leukaemia cells?
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DOI：
10.1016/j.phrs.2012.10.012
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发表时间：
2013-01-01
影响因子：
9.3
通讯作者：
Mlejnek, Petr
中科院分区：
文献类型：
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作者：
Kosztyu, Petr;Dolezel, Petr;Mlejnek, Petr
研究P-糖蛋白(P-gp、ABCB1、MDR1)在人白血病细胞对尼洛替尼耐药中的作用。我们使用高表达P-gp的K562/Dox细胞及其变异体(亚克隆)，并逐渐降低P-gp的表达。用针对ABCB1基因的shRNA表达载体稳定地转染K562/Dox细胞，获得了这些亚克隆。用特定的荧光探针分析P-gp的功能表明，染料外流逐渐减少，这与P-gp的表达成正比。我们观察到，与不表达P-gp的K562/Dox细胞相比，高表达P-gp的K562/Dox细胞内尼洛替尼的水平显著降低。伴随这一效应的是K562/Dox细胞对尼洛替尼的敏感性降低。重要的是，P-gp表达下调的细胞逐渐失去了降低细胞内尼洛替尼水平的能力，尽管它们仍然显著降低了柔红霉素(DNR)的细胞内水平。相应地，P-gp表达降低的细胞不能对尼洛替尼产生耐药性，但它们对柔红霉素表现出明显的耐药性。我们证明了P-gp是否参与尼洛替尼耐药的结论强烈依赖于其在蛋白水平的表达。(C)爱思唯尔有限公司出版的2012年。
The effect of P-glycoprotein (P-gp, ABCB1, MDR1) expression on cell resistance to nilotinib was studied in human leukaemia cells. We used K562/Dox cells overexpressing P-gp and their variants (subclones) with a gradually decreased P-gp expression. These subclones were established by stable transfection of K562/Dox cells with a plasmid vector expressing shRNA targeting the ABCB1 gene. Functional analysis of P-gp using a specific fluorescent probe indicated gradually decreased dye efflux which was proportional to the P-gp expression. We observed that K562/Dox cells overexpressing P-gp contained a significantly reduced intracellular level of nilotinib when compared to their counter partner K562 cells, which do not express P-gp. This effect was accompanied by a decreased sensitivity of the K562/Dox cells to nilotinib. Importantly, cells with downregulated expression of P-gp gradually lost their ability to decrease the intracellular level of nilotinib although they still significantly decreased the intracellular level of daunorubicin (DNR). Accordingly, cells with the reduced expression of P-gp concomitantly failed to provide resistance to nilotinib, however, they exhibited a significant resistance to DNR Taken together, we demonstrated that the conclusion as to whether P-gp is involved in nilotinib resistance or not strongly depends on its expression at protein level. (c) 2012 Published by Elsevier Ltd.