Wnt/β-catenin pathway regulates ABCB1 transcription in chronic myeloid leukemia.
Wnt/β-catenin pathway regulates ABCB1 transcription in chronic myeloid leukemia.
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DOI：
10.1186/1471-2407-12-303
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发表时间：
2012-07-23
期刊：
影响因子：
3.8
通讯作者：
Abdelhay E
中科院分区：
文献类型：
--
作者：
Corrêa S;Binato R;Du Rocher B;Castelo-Branco MT;Pizzatti L;Abdelhay E
众所周知，慢性粒细胞白血病(CML)的晚期患者对治疗更具抵抗力。这种耐药性与ABCB1的过度表达有关，ABCB1的过度表达导致了多药耐药(MDR)现象。MDR的特点是对非相关药物耐药，P糖蛋白(由ABCB1编码)被认为是其出现的主要原因。WNT信号已被证明在慢性粒细胞白血病的几个方面都很重要。最近，在其他类型的癌症中，Wnt信号通过其典型的通路与ABCB1的调控联系在一起，该通路由β-连环蛋白介导。在本研究中，我们研究了Wnt/β-catenin通路在慢性粒细胞白血病中的转录调控作用，因为ABCB1的基本启动子有几个β-catenin结合位点。β-连环蛋白是典型的Wnt信号的中介体，在慢性粒细胞白血病的进展中起重要作用。在本工作中，我们以K562细胞系及其衍生的耐多药细胞系Lucena(K562/VCR)为研究模型。采用实时定量聚合酶链式反应(RT-qPCR)、凝胶迁移率改变分析(EMSA)、染色质免疫沉淀(ChIP)、流式细胞仪(FACS)、免疫印迹、免疫荧光、RNA敲除(SiRNA)和荧光素酶报告等方法。β-连环蛋白在体外存在于两种细胞系的ABCB1启动子蛋白复合体中，但在体内耐药细胞系中其结合更为明显。与其亲本细胞系相比，卢塞纳细胞也表现出更高的β-连环蛋白水平。Wnt1和β-catenin的缺失和核β-catenin的过度表达以及TcF结合位点的激活表明，Wnt信号的典型途径正向调节了ABCB1的表达。这些结果首次表明，Wnt/β-catenin途径调节慢性粒细胞白血病的ABCB1。
The advanced phases of chronic myeloid leukemia (CML) are known to be more resistant to therapy. This resistance has been associated with the overexpression of ABCB1, which gives rise to the multidrug resistance (MDR) phenomenon. MDR is characterized by resistance to nonrelated drugs, and P-glycoprotein (encoded by ABCB1) has been implicated as the major cause of its emergence. Wnt signaling has been demonstrated to be important in several aspects of CML. Recently, Wnt signaling was linked to ABCB1 regulation through its canonical pathway, which is mediated by β-catenin, in other types of cancer. In this study, we investigated the involvement of the Wnt/β-catenin pathway in the regulation of ABCB1 transcription in CML, as the basal promoter of ABCB1 has several β-catenin binding sites. β-catenin is the mediator of canonical Wnt signaling, which is important for CML progression. In this work we used the K562 cell line and its derived MDR-resistant cell line Lucena (K562/VCR) as CML study models. Real time PCR (RT-qPCR), electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP), flow cytometry (FACS), western blot, immunofluorescence, RNA knockdown (siRNA) and Luciferase reporter approaches were used. β-catenin was present in the protein complex on the basal promoter of ABCB1 in both cell lines in vitro, but its binding was more pronounced in the resistant cell line in vivo. Lucena cells also exhibited higher β-catenin levels compared to its parental cell line. Wnt1 and β-catenin depletion and overexpression of nuclear β-catenin, together with TCF binding sites activation demonstrated that ABCB1 is positively regulated by the canonical pathway of Wnt signaling. These results suggest, for the first time, that the Wnt/β-catenin pathway regulates ABCB1 in CML.
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影响因子：
50.3
作者：
Gregory MA;Phang TL;Neviani P;Alvarez-Calderon F;Eide CA;O'Hare T;Zaberezhnyy V;Williams RT;Druker BJ;Perrotti D;Degregori J
通讯作者：
Degregori J
影响因子：
20.3
作者：
Mahon, FX;Belloc, F;Melo, JV
通讯作者：
Melo, JV
影响因子：
158.5
作者：
Jamieson, CHM;Ailles, LE;Weissman, IL
通讯作者：
Weissman, IL
影响因子：
6.7
作者：
Gurney, H.;Wong, M.;Schran, H.
通讯作者：
Schran, H.
影响因子：
11.4
作者：
Hu, Y.;Chen, Y.;Li, S.
通讯作者：
Li, S.