AQP5 promotes epithelial-mesenchymal transition and tumor growth through activating the Wnt/β-catenin pathway in triple-negative breast cancer

IF 1.5 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis Pub Date : 2024-07-01 DOI:10.1016/j.mrfmmm.2024.111868

Zhengcai Zhu, Tao Li, Honggang Wang, Lianghe Jiao

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引用次数: 0

Abstract

Background

Emerging data identifies aquaporin 5 (AQP5) as a vital player in many kinds of cancers. Over expression of AQP5 was associated with increased metastasis and poor prognosis, suggesting that AQP5 may facilitate cancer cell proliferation and migration. Our previous studies also showed that AQP3 and AQP5 were highly expressed in triple-negative breast cancer (TNBC) and the expression of AQP3 and AQP5 in TNBC tissue was positive correlated with advanced clinical stage.

Objective

We aim to investigate the role of AQP5 in TNBC oncogenesis and development.

Methods

MDA-MB-231 cells were transfected with siRNA-AQP5 and AQP5 overexpression vector to establish a differential expression system for AQP5. Cell proliferation and apoptosis of MDA-MB-231 cells were detected by CCK-8 (Cell Counting Kit-8) and FCM (flow cytometry), respectively. Cell migration and invasion abilities were evaluated by wound healing assay and transwell assay. The qRT-PCR and western blot assays were used to study the effect of AQP5 expression level on the expression of epithelial-to-mesenchymal transition (EMT) related molecules. The effects of ICG-001, a Wnt/β-catenin signaling pathway inhibitor, on the invasive and migratory capabilities of overexpressed AQP5 cells and downstream molecules were measured.

Results

1. The expression of AQP5 in the MDA-MB-231 cells was significantly higher than that in the MCF-10A cells. 2. Up-regulation of AQP5 significantly promoted the proliferation, migration and invasion of TNBC cells, while inhibited the cell apoptosis; in addition, up-regulation of AQP5 increased the expression of Bcl-2 and decreased the expression of Caspase-3. However, knockdown of AQP5 presented the adverse effects of AQP5 overexpression. 3. Overexpressed AQP5 induced the overexpression of EMT-related factors, which further promoted the migration and invasion of cells. 4. Overexpression of AQP5 could up-regulate the expression of β-catenin in the nucleus followed by increasing the expression levels of downstream genes in Wnt/β-catenin signaling pathway. Moreover, ICG-001, the inhibitor of Wnt/β-catenin signaling pathway, could significantly attenuate the effect of overexpression of AQP5 on cells, further confirming that AQP5 may promote the proliferation, migration and invasion of TNBC cells by activating Wnt/β-catenin signaling pathway.

Conclusions

In the TNBC cells, AQP5 modulates the expression levels of EMT-related proteins through activation of Wnt/β-catenin signaling pathway, thus enhancing the cell proliferation, migration and invasion while inhibiting the cell apoptosis.

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AQP5通过激活三阴性乳腺癌中的Wnt/β-catenin通路促进上皮-间质转化和肿瘤生长

背景：越来越多的数据表明，水蒸气素 5（AQP5）在多种癌症中发挥着重要作用。AQP5 的过度表达与转移增加和预后不良有关，这表明 AQP5 可能会促进癌细胞的增殖和迁移。我们之前的研究也表明，AQP3 和 AQP5 在三阴性乳腺癌（TNBC）中高表达，且 AQP3 和 AQP5 在 TNBC 组织中的表达与晚期临床分期呈正相关：我们旨在研究 AQP5 在 TNBC 肿瘤发生和发展过程中的作用：方法：用 siRNA-AQP5 和 AQP5 过表达载体转染 MDA-MB-231 细胞，建立 AQP5 差异表达系统。分别用 CCK-8（细胞计数试剂盒-8）和 FCM（流式细胞术）检测 MDA-MB-231 细胞的增殖和凋亡。细胞迁移和侵袭能力通过伤口愈合试验和透孔试验进行评估。qRT-PCR 和 Western 印迹检测用于研究 AQP5 表达水平对上皮细胞向间质转化（EMT）相关分子表达的影响。还测定了Wnt/β-catenin信号通路抑制剂ICG-001对过表达AQP5细胞的侵袭和迁移能力及下游分子的影响：1.1. AQP5在MDA-MB-231细胞中的表达量明显高于MCF-10A细胞。2.2. AQP5的上调能明显促进TNBC细胞的增殖、迁移和侵袭，同时抑制细胞凋亡；此外，AQP5的上调能增加Bcl-2的表达，降低Caspase-3的表达。然而，敲除AQP5会出现AQP5过表达的不良反应。3.过表达的 AQP5 会诱导 EMT 相关因子的过表达，从而进一步促进细胞的迁移和侵袭。4.4. AQP5的过表达可上调细胞核中β-catenin的表达，进而增加Wnt/β-catenin信号通路下游基因的表达水平。此外，Wnt/β-catenin信号通路抑制剂ICG-001能显著减弱AQP5过表达对细胞的影响，进一步证实了AQP5可通过激活Wnt/β-catenin信号通路促进TNBC细胞的增殖、迁移和侵袭：结论：在 TNBC 细胞中，AQP5 通过激活 Wnt/β-catenin 信号通路调节 EMT 相关蛋白的表达水平，从而在抑制细胞凋亡的同时促进细胞增殖、迁移和侵袭。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Mutation Research-Fundamental and Molecular Mechanisms of Mutagenesis 生物-毒理学

CiteScore

4.90

自引率

0.00%

发文量

审稿时长

51 days

期刊介绍： Mutation Research (MR) provides a platform for publishing all aspects of DNA mutations and epimutations, from basic evolutionary aspects to translational applications in genetic and epigenetic diagnostics and therapy. Mutations are defined as all possible alterations in DNA sequence and sequence organization, from point mutations to genome structural variation, chromosomal aberrations and aneuploidy. Epimutations are defined as alterations in the epigenome, i.e., changes in DNA methylation, histone modification and small regulatory RNAs. MR publishes articles in the following areas: Of special interest are basic mechanisms through which DNA damage and mutations impact development and differentiation, stem cell biology and cell fate in general, including various forms of cell death and cellular senescence. The study of genome instability in human molecular epidemiology and in relation to complex phenotypes, such as human disease, is considered a growing area of importance. Mechanisms of (epi)mutation induction, for example, during DNA repair, replication or recombination; novel methods of (epi)mutation detection, with a focus on ultra-high-throughput sequencing. Landscape of somatic mutations and epimutations in cancer and aging. Role of de novo mutations in human disease and aging; mutations in population genomics. Interactions between mutations and epimutations. The role of epimutations in chromatin structure and function. Mitochondrial DNA mutations and their consequences in terms of human disease and aging. Novel ways to generate mutations and epimutations in cell lines and animal models.