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Gene expression profiling of Lentivirus-mediated Gene Silencing of RARβ Inhibits A549 Non-Small Cell Lung Cancer stem cells

ABSTRACT: Retinoic acid Receptor Beta (RARβ) protein encoded by RARβ gene is a nuclear receptor protein that binds to retinoic acid (RA) to mediate RA function in cellular signalling in embryogenic morphogenesis, cell growth and differentiation. However, the function of RARβ in cancer stem cells (CSCs) maintenance of non-small cell lung cancer (NSCLC) is yet to be determined We used microarrays to analyse the trancriptomic changes in lung cancer and cancer stem cells following RARβ silencing.

ORGANISM(S): Homo sapiens

PROVIDER: GSE134788 | GEO | 2020/07/24

REPOSITORIES: GEO

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Project description:Microarray analysis reveals up-regulation of retinoic acid and hepatocyte growth factor related signaling pathways by pro-insulin C-peptide in kidney proximal tubular cells: Antagonism of the pro-fibrotic effects of TGF-b1 Novel signaling roles for C-peptide have recently been discovered with evidence that it can ameliorate complications of type 1 diabetes. Here we sought to identify new pathways regulated by C-peptide of relevance to the pathophysiology of diabetic nephropathy. Microarray analysis was performed to identify genes regulated by either C-peptide and/or transforming growth factor beta 1 (TGF-β1) in a human proximal tubular cell line, HK-2. Expression of retinoic acid receptor β (RARβ), hepatcoyte growth factor (HGF), cellular retinoic acid binding protein II (CRABPII), vimentin, E-cadherin, Snail and β-catenin was assessed by immunoblotting. The cellular localisation of vimentin and β-catenin was determined by immunocytochemistry. Changes in cell morphology were assessed by phase contrast microscopy. Gene expression profiling demonstrated differential expression of 953 and 1,458 genes after C-peptide exposure for 18h or 48h respectively. From these, members of the anti-fibrotic retinoic acid (RA) and HGF signaling pathways were selected. Immunoblotting demonstrated that C-peptide increased RARβ, CRABPII and HGF. We confirmed a role for RA in reversal of TGF-β1-induced changes associated with epithelial-mesenchymal transition (EMT), including expression changes in Snail, E-cadherin, vimetin and redistribution of β-catenin. Importantly, these TGF-β1-induced changes were inhibited by C-peptide. Further, effects of TGF-β1 on Snail and E-cadherin expression were blocked by HGF and inhibitory effects of C-peptide were removed by blockade of HGF activity. This study identifies a novel role for HGF as an effector of C-peptide, possibly via an RA signaling pathway, highlighting C-peptide as a potential therapy for diabetic nephropathy.

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Gene expression profiling of Lentivirus-mediated Gene Silencing of RARβ Inhibits A549 Non-Small Cell Lung Cancer stem cells

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