NF-κB Participates in the Stem Cell Phenotype of Ovarian Cancer Cells
NF-κB Participates in the Stem Cell Phenotype of Ovarian Cancer Cells
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DOI：
10.1016/j.arcmed.2017.08.001
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发表时间：
2017-05-01
影响因子：
7.7
通讯作者：
Maldonado, Vilma
中科院分区：
文献类型：
--
作者：
Gonzalez-Torres, Carolina;Gaytan-Cervantes, Javier;Maldonado, Vilma
背景。nf - κ B是一种参与肿瘤干细胞维持的转录因子。对于卵巢癌中这一途径的特异性茎相关上游调控因子知之甚少。本研究的目的是分析nf - κ B信号通路在卵巢癌干细胞中的作用。采用分选细胞术分离干细胞。Western blot和RT-PCR检测蛋白和信使RNA水平。分别使用sirna和显性阴性蛋白进行功能损失和功能获得分析。用报告基因法测定NF-kappa B结合活性。用软琼脂、菌落形成、卵球形成和体内致瘤性测定来估计茎的表型。SKOV3卵巢癌细胞系CD44+亚群的关键严厉性基因mRNA水平升高，致瘤能力增强，RelA、RelB和IKK α表达增加。当典型途径被IkB α的显性阴性版本抑制时，干细胞群减少，如CD44+亚群减少，严厉基因表达减少和茎表型减少所示。此外，主要的上游非典型激酶IKK α在CSC群体中高表达。因此，当使用shrna抑制IKK α时，严厉基因的表达减少。该报告首次显示NF-kappa B通路的几个要素在维持卵巢癌干细胞群中的重要性。(c) 2017 imss。Elsevier Inc.出版。
Background. NF-kappa B is a transcription factor involved in cancer stem cells maintenance of many tumors. Little is known about the specific stem-associated upstream regulators of this pathway in ovarian cancer. The Aim of the study was to analyze the role of the canonical and non-canonical NF-kappa B pathways in stem cells of ovarian cancer cell lines.Methods. Stem cells were isolated using sorting cytometry. Western blot and RT-PCR were used to quantify protein and messenger RNA levels. Loss and gain of function assays were performed using siRNAs and dominant-negative proteins, respectively. NF-kappa B binding activity was measured with a reporter gene assay. The stem phenotype was estimated with clonogenic assays using soft agar, colony formation, ovospheres formation and in vivo tumorigenicity assays.Results. The CD44+ subpopulation of SKOV3 ovarian cancer cell line presented higher mRNA levels of key sternness genes, an increased tumorigenic capacity and higher expression of the RelA, RelB and IKK alpha. When the canonical pathway was inhibited by means of a dominant-negative version of IkB alpha, the stem cell population was reduced, as shown by a reduced CD44+ subpopulation, a decrease in the expression of the sternness genes and a reduction of the stem phenotype. In addition, IKK alpha, the main upstream non-canonical kinase, was highly expressed in the CSC population. Accordingly, when IKK alpha was inhibited using shRNAs, the expression of the sternness genes was reduced.Conclusions. This report is the first to show the importance of several elements of both NF-kappa B pathway in maintaining the ovarian cancer stem cell population. (C) 2017 IMSS. Published by Elsevier Inc.