The Association of RGS2 and Slug in the Androgen-induced Acquisition of Mesenchymal Features of Breast MDA-MB-453 Cancer Cells.

IF 1.5 4区 医学 Q4 ENDOCRINOLOGY & METABOLISM
Endocrine Research Pub Date : 2022-02-01 Epub Date: 2022-02-16 DOI:10.1080/07435800.2022.2036752
Dana B Alsafadi, Mohammad S Abdullah, Randa Bawadi, Mamoun Ahram
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引用次数: 5

Abstract

Background: Epithelial-mesenchymal transition (EMT) of tumor cells is a prerequisite to cancer cell invasion and metastasis. This process involves a network of molecular alterations. Androgen receptor (AR) plays an important role in the biology of breast cancers, particularly those dependent on AR expression like luminal AR (LAR) breast cancer subtype. We have recently reported that the AR agonist, dihydrotestosterone (DHT), induces a mesenchymal transition of MDA-MB-453 cells, concomitant with transcriptional up-regulation of Slug and regulator of G protein signaling 2 (RGS2).

Objective: The role of Slug and RGS2 in mediating the DHT-induced effects in these cells was investigated.

Methods: MDA-MB-453 cells were used as a model system of LAR breast cancer. Immunofluorescence was used to examine cell morphology and protein localization. Protein expression was analyzed by immunoblotting. Protein localization was confirmed by cell fractionation followed by immunoblotting. Protein-protein interaction was confirmed by co-immunoprecipitation followed by immunoblotting. Transwell membranes were used to assess cell migration. Transfection of cells with siRNA molecules that target Slug and RGS2 mRNA was utilized to delineate the modes of action of these two molecules.

Results: Treatment of MDA-MB-453 cells with DHT induced the expression of both proteins. In addition, AR-Slug, AR-RGS2, and Slug-RGS2 interactions were observed shortly after AR activation. Knocking down Slug abrogated the basal, but not the DHT-induced, cell migration and blocked DHT-induced mesenchymal transition. On the other hand, RGS2 knocked-down cells had an increased level of Slug protein and assumed mesenchymal cell morphology with induced migration, and the addition of DHT further elongated cell morphology and stimulated their migration. Inhibition of AR or β-catenin reverted the RGS2 knocked-down cells to the epithelial phenotype, but only inhibition of AR blocked their DHT-induced migration.

Conclusions: These results suggest the involvement of RGS2 and Slug in a complex molecular network regulating the DHT-induced mesenchymal features in MDA-MB-453 cells. The study may offer a better understanding of the biological role of AR in breast cancer toward devising AR-based therapeutic strategies.

RGS2和Slug在雄激素诱导的乳腺MDA-MB-453癌细胞间充质特征获取中的关联
背景:肿瘤细胞的上皮-间质转化(Epithelial-mesenchymal transition, EMT)是癌细胞侵袭和转移的先决条件。这个过程涉及到一个分子变化网络。雄激素受体(AR)在乳腺癌的生物学中起着重要作用,特别是那些依赖于AR表达的乳腺癌,如腔内AR (LAR)亚型。我们最近报道了AR激动剂双氢睾酮(DHT)诱导MDA-MB-453细胞的间质转化,同时伴有Slug的转录上调和G蛋白信号传导2 (RGS2)的调节。目的:探讨Slug和RGS2在dht诱导的细胞中的作用。方法:采用MDA-MB-453细胞作为LAR乳腺癌模型系统。免疫荧光法检测细胞形态和蛋白定位。免疫印迹法分析蛋白表达。通过细胞分离和免疫印迹确认蛋白定位。通过免疫共沉淀法和免疫印迹法确认蛋白-蛋白相互作用。Transwell膜用于评估细胞迁移。用靶向Slug和RGS2 mRNA的siRNA分子转染细胞来描述这两种分子的作用模式。结果:用DHT处理MDA-MB-453细胞可诱导这两种蛋白的表达。此外,AR激活后不久,AR- slug、AR- rgs2和Slug-RGS2相互作用被观察到。敲除Slug可消除dht诱导的基础细胞迁移,而非dht诱导的细胞迁移,并可阻断dht诱导的间质转化。另一方面,RGS2敲低的细胞Slug蛋白水平升高,呈现间充质细胞形态并诱导迁移,DHT的加入进一步拉长了细胞形态并刺激其迁移。抑制AR或β-catenin可使RGS2敲除的细胞恢复到上皮表型,但只有抑制AR才能阻断dht诱导的迁移。结论:这些结果表明RGS2和Slug参与调控dht诱导的MDA-MB-453细胞间充质特征的复杂分子网络。该研究可能有助于更好地了解AR在乳腺癌中的生物学作用,从而制定基于AR的治疗策略。
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来源期刊
Endocrine Research
Endocrine Research 医学-内分泌学与代谢
CiteScore
4.30
自引率
0.00%
发文量
10
审稿时长
>12 weeks
期刊介绍: This journal publishes original articles relating to endocrinology in the broadest context. Subjects of interest include: receptors and mechanism of action of hormones, methodological advances in the detection and measurement of hormones; structure and chemical properties of hormones. Invitations to submit Brief Reviews are issued to specific authors by the Editors.
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