Effects of patient pleural effusion fluids on the BBSome components expression of human benign mesothelial cells.

IF 3.6 2区 医学 Q1 PHYSIOLOGY
Rajesh M Jagirdar, Erasmia Rouka, Eleanna Pitaraki, Ioannis Sarrigeorgiou, Ourania S Kotsiou, Sotirios I Sinis, Eleftherios D Papazoglou, Periklis Marnas, Zoi Malami, Peggy Lymberi, Anastasios D Giannou, Chrissi Hatzoglou, Konstantinos I Gourgoulianis, Sotirios G Zarogiannis
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Abstract

Background: Malignant pleural mesothelial cells are affected by the extracellular milieu while such data on benign cells are scarce. Benign cells sense the extracellular environment with the Primary Cilium (PC) and its molecular complex, the BBSome, is critical for this process. Here we aimed at assessing the changes in BBSome genes expression in ordinary 2D and spheroid 3D cell cultures after incubation with pleural effusion fluids (PF) of several etiologies.

Methods: Benign human mesothelial cells MeT-5A were incubated with PF from patients with mesothelioma (Meso-PF), breast cancer (BrCa-PF), hemothorax (Hemo-PF) and congestive heart failure (CHF-PF). Gene expression of BBS1, 2, 4, 5, 7, 9, 18 was assessed by quantitative real-time PCR (qRT-PCR) to monitor PF-induced gene expression changes. MeT-5A cell migration using the PC-modulating drugs ammonium sulfate (AS) and lithium chloride (LC) during PF incubation was also determined.

Results: BBSome gene expression upon influence of BrCa-PF and Hemo-PF was more pronounced in 2D compared to 3D, inducing global changes in 2D. CHF-PF and Meso-PF also induced changes in 2D but not as many, while in all cases MeT-5A grown in 3D were more resistant to the effects of the PF. Meso-PF decreased 2D cell migration, while the disturbance of PC in all PF cases resulted in decreased cell migration.

Conclusions: These data suggest distinct BBSome molecular profile changes in benign mesothelial cells exposed to malignant and benign PF, in each case, in both 2D and 3D. Cell migration is sensitive to drug disturbance with PC modulators in PF-exposed cells.

患者胸腔积液对人类良性间皮细胞 BBSome 成分表达的影响
背景:恶性胸膜间皮细胞会受到细胞外环境的影响,而良性细胞的此类数据却很少。良性细胞通过初级纤毛(PC)感知细胞外环境,其分子复合物 BBSome 对这一过程至关重要。在此,我们旨在评估普通二维和球形三维细胞培养物在与多种病因的胸腔积液(PF)培养后 BBSome 基因表达的变化:方法:将良性人间皮细胞 MeT-5A 与间皮瘤(Meso-PF)、乳腺癌(BrCa-PF)、血胸(Hemo-PF)和充血性心力衰竭(CHF-PF)患者的胸腔积液培养。通过实时定量 PCR(qRT-PCR)评估 BBS1、2、4、5、7、9、18 的基因表达,监测 PF 诱导的基因表达变化。此外,还利用 PC 调节药物硫酸铵(AS)和氯化锂(LC)测定了 PF 培养期间 MeT-5A 细胞的迁移情况:结果:BrCa-PF 和 Hemo-PF 影响下的 BBSome 基因表达在 2D 中比在 3D 中更明显,在 2D 中引起了整体变化。CHF-PF和Meso-PF也诱导二维细胞发生变化,但变化幅度较小,而在所有情况下,三维生长的MeT-5A对PF的影响更有抵抗力。Meso-PF减少了二维细胞的迁移,而所有PF情况下PC的干扰都导致细胞迁移减少:这些数据表明,暴露于恶性和良性间皮细胞的良性间皮细胞在二维和三维情况下都会发生不同的 BBSome 分子谱变化。在暴露于 PF 的细胞中,细胞迁移对 PC 调节剂的药物干扰很敏感。
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来源期刊
CiteScore
9.20
自引率
4.10%
发文量
146
审稿时长
2 months
期刊介绍: The American Journal of Physiology-Lung Cellular and Molecular Physiology publishes original research covering the broad scope of molecular, cellular, and integrative aspects of normal and abnormal function of cells and components of the respiratory system. Areas of interest include conducting airways, pulmonary circulation, lung endothelial and epithelial cells, the pleura, neuroendocrine and immunologic cells in the lung, neural cells involved in control of breathing, and cells of the diaphragm and thoracic muscles. The processes to be covered in the Journal include gas-exchange, metabolic control at the cellular level, intracellular signaling, gene expression, genomics, macromolecules and their turnover, cell-cell and cell-matrix interactions, cell motility, secretory mechanisms, membrane function, surfactant, matrix components, mucus and lining materials, lung defenses, macrophage function, transport of salt, water and protein, development and differentiation of the respiratory system, and response to the environment.
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