Long noncoding RNA expression analysis reveals the regulatory effects of nitinol-based nanotubular coatings on human coronary artery endothelial cells

Abstract

Pan Shang,1,* Gan Chen,1,* Guannan Zu,2 Xiang Song,1 Peng Jiao,2 Guoxing You,1 Jingxiang Zhao,1 Hongyi Li,2 Hong Zhou11Academy of Military Medical Sciences, Institute of Health Service and Transfusion Medicine, Beijing 100850, People’s Republic of China; 2Photoelectrochemical Research Group, School of Materials Science and Engineering, Beijing University of Technology, Beijing 100124, People’s Republic of China *These authors contributed equally to this workBackground: Cardiovascular disease (CVD) is the leading cause of mortality all over the world. Vascular stents are used to ameliorate vascular stenosis and recover vascular function. The application of nanotubular coatings has been confirmed to promote endothelial cell (EC) proliferation and function. However, the regulatory mechanisms involved in cellular responses to the nanotubular topography have not been defined. In the present study, a microarray analysis was performed to explore the expression patterns of long noncoding RNAs (lncRNAs) in human coronary artery endothelial cells (HCAECs) that were differentially expressed in response to nitinol-based nanotubular coatings.Materials and methods: First, anodization was performed to synthesize nitinol-based nanotubular coatings. Then, HCAECs were cultured on the samples for 24 h to evaluate cell cytoskeleton organization. Next, total RNA was extracted and synthesized into cRNA, which was hybridized onto the microarray. GO analysis and KEGG pathway analysis were performed to investigate the roles of differentially expressed messenger RNAs (mRNAs). Quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR) was performed to validate the expression of randomly selected lncRNAs. Coexpression networks were created to identify the interactions among lncRNAs and the protein-coding genes involved in nanotubular topography-induced biological and molecular pathways. Independent Student’s t-test was applied for comparisons between two groups with statistical significance set at p<0.05.Results: 1085 lncRNAs and 227 mRNAs were significantly differentially expressed in the nitinol-based nanotubular coating group. Bioinformatics analysis revealed that extracellular matrix receptor interactions and cell adhesion molecules play critical roles in the sensing of nitinol-based nanotubular coatings by HCAECs. The TATA-binding protein (TBP) and TBP-associated transfactor 1 (TAF1) are important molecules in EC responses to substrate topography.Conclusion: This study suggests that nanotubular substrate topography regulates ECs by differentially expressed lncRNAs involved extracellular matrix receptor interactions and cell adhesion molecules.Keywords: nitinol titanium dioxide nanotubes, RNA sequencing, long noncoding RNA, nanotopography, molecular networks

Keywords

• Nitinol titanium dioxide nanotubes
• RNA sequencing
• Long noncoding RNA
• Nanotopography
• Molecular networks