Involvement of CUL4A in regulation of multidrug resistance to P-gp substrate drugs in breast cancer cells.
Involvement of CUL4A in regulation of multidrug resistance to P-gp substrate drugs in breast cancer cells.
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CUL4A 参与乳腺癌细胞对 P-gp 底物药物多药耐药性的调节
DOI：
10.3390/molecules19010159
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发表时间：
2013-12-24
期刊：
影响因子：
--
通讯作者：
Wei G
中科院分区：
文献类型：
--
作者：
Wang Y;Ma G;Wang Q;Wen M;Xu Y;He X;Zhang P;Wang Y;Yang T;Zhan P;Wei G
CUL4A编码基于cullin的E3泛素连接酶复合体的核心成分，该复合体通过靶向多种蛋白质的泛素化和降解来调节许多关键过程，如细胞周期进程、DNA复制、DNA修复和染色质重塑。在这份报告中描述的研究中，我们旨在阐明CUL4A是否参与了乳腺癌细胞的多药耐药(MDR)。我们首先将携带CUL4A和特异性shCUL4A的载体分别导入乳腺癌细胞和相应的ADR细胞。通过逆转录聚合酶链式反应和Western blotting，我们发现CUL4A在mCF7和mda-MB-468细胞中的过表达在转录和蛋白水平上都上调了mdr1/P-gp的表达，从而导致了对P-gp底物药物的多药耐药，这是通过四甲基偶氮唑蓝(3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)法检测到的。另一方面，沉默MCF7/ADR和MDA-MB-468/ADR细胞中的CUL4A则产生相反的效果。此外，CUL4A过表达细胞中的ERK1/2被高度激活，经ERK1/2特异性抑制剂PD98059处理后，CUL4A诱导的MDR1/P-gp表达显著降低。免疫组织化学结果显示，乳腺癌组织中P-gp的表达与CUL4A和ERK1/2的表达呈正相关，提示CUL4A和ERK1/2通过调控MDR1/P-gp的表达参与乳腺癌的多药耐药。
CUL4A encodes a core component of a cullin-based E3 ubiquitin ligase complex that regulates many critical processes such as cell cycle progression, DNA replication, DNA repair and chromatin remodeling by targeting a variety of proteins for ubiquitination and degradation. In the research described in this report we aimed to clarify whether CUL4A participates in multiple drug resistance (MDR) in breast cancer cells. We first transfected vectors carrying CUL4A and specific shCUL4A into breast cancer cells and corresponding Adr cells respectively. Using reverse transcription polymerase chain reactions and western blots, we found that overexpression of CUL4A in MCF7 and MDA-MB-468 cells up-regulated MDR1/P-gp expression on both the transcription and protein levels, which conferred multidrug resistance to P-gp substrate drugs, as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. On the other hand, silencing CUL4A in MCF7/Adr and MDA-MB-468/Adr cells led to the opposite effect. Moreover, ERK1/2 in CUL4A-overexpressing cells was highly activated and after treatment with PD98059, an ERK1/2-specific inhibitor, CUL4A-induced expression of MDR1/P-gp was decreased significantly. Lastly, immunohistochemistry in breast cancer tissues showed that P-gp expression had a positive correlation with the expression of CUL4A and ERK1/2. Thus, these results implied that CUL4A and ERK1/2 participated in multi-drug resistance in breast cancer through regulation of MDR1/P-gp expression.
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