Malignant pleural mesothelial cells are affected by the extracellular milieu although such data on benign cells are scarce. Benign cells sense the extracellular environment with the primary cilium (PC) and its molecular complex, the Bardet-Biedl syndrome family of proteins (BBSome), is critical for this process. Here we aimed to assess the changes in BBSome gene expression in ordinary two-dimensional (2-D) and spheroid three-dimensional (3-D) cell cultures after incubation with pleural effusion fluids (PFs) of several etiologies. The 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, and 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. BBSome gene expression upon influence of BrCa-PF and Hemo-PF was more pronounced in 2-D compared with 3-D, inducing global changes in 2-D. CHF-PF and Meso-PF also induced changes in 2-D but not as many, while in all cases, MeT-5A grown in 3-D were more resistant to the effects of the PF. Meso-PF decreased 2-D cell migration, while the disturbance of PC in all PF cases resulted in decreased cell migration. These data suggest distinct BBSome molecular profile changes in benign mesothelial cells exposed to malignant and benign PF that is different in each case, in both 2-D and 3-D. Cell migration is sensitive to drug disturbance with PC modulators in PF-exposed cells.NEW & NOTEWORTHY Studying mesothelial PC in pleural physiology and pathophysiology is at an early stage. Previously, we have highlighted the role of the PC in mesothelial cell phenotypes as well as the role of BBSome components in the context of benign and malignant mesothelial cell physiology. Here we extended our contributions by providing evidence on the BBSome changes induced in benign mesothelial cells by their exposure to different etiology PFs.
Keywords: 3-D culture; BBSome; malignant pleural mesothelioma; mesothelial cells; pleural effusion.