ERS Respiratory Channel

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Abstract

Introduction:Primary Cilium (PC) is an apical sensory organelle receiving extracellular cues such as inflammatory ones. The PC is composed of Bardet-Biedl family of proteins that form the BBSome complex. Mesothelial cells lining the pleural cavity have to be described for PC components. Pleural effusion formation is accompanied by inflammation, with uknown impact on BBSome expression. We assessed BBsome gene(s) expression from inflammatory stress with bacterial lipopolysaccharide (LPS).

Materials and methods: Benign mesothelial cells, MeT-5A, and mesothelioma primary cultures established from a patient with epithelioid mesothelioma (pMPM) were cultured with regular medium or media with 5?g/ml of LPS for 24 hours. Cells were lysed for RNA processing and cDNA was prepared followed by qPCR quantitation with gene specific primers for BBS1, BBS2, BBS4, BBS5, BBS7, BBS9 and BBS18 and ?actin. Log₂ transformed 2^-??Ct values were used to compare relative gene expression.

Results:In MeT-5A cells assayed BBS genes? expression was detectable but without significant differences and no BBS5 expression. In malignant pMPM cells, BBS9 was significantly downregulated (Con mean±SEM;0.0±0.11, LPS -1.11±0.32,p<0.01). BBS5 was expressed in pMPM but not regulated.

Conclusion:Benign cells maintained steady BBS genes? expression during LPS stimulus. The expression patterns in the assessed BBSome genes are reversed when compared between benign and neoplastic cells. BBS9 in primary mesothelioma cells is downregulated from LPS stimulus. The findings on BBSome gene expression patterns and differential of BBS9 due to LPS stimulus in transformed cells warrants further investigation.

Abstract

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