RESUMO
BACKGROUND: Although long non-coding RNAs (lncRNAs) have been reported to serve as potential biomarkers of atherosclerosis (AS), the role of lncRNA small nucleolar RNA host gene 12 (SNHG12) in AS still remains to be elucidated. OBJECTIVES: The present study aimed to investigate the regulatory effects and potential mechanisms of SNHG12 in human vascular smooth muscle cells (hVSMCs). MATERIAL AND METHODS: Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) was employed to determine the expression of SNHG12, miR-766-5p and eukaryotic translation initiation factor 5A (EIF5A) in oxidized low-density lipoprotein (ox-LDL)-induced hVSMCs. After transfection with short hairpin RNA (shRNA)-SNHG12, cell viability was estimated using the Cell Counting Kit-8 (CCK-8) assay. Wound healing and transwell assays were used for evaluating migratory capacities of hVSMCs. To further investigate the regulatory mechanisms, binding sites between SNHG12 and miR-766-5p, and EIF5A and miR-766-5p were predicted using starBase database and validated using luciferase reporter gene assays. Moreover, cell viability and migration were detected following EIF5A overexpression and SNHG12-knockdown. RESULTS: SNHG12 was significantly upregulated in ox-LDL-induced hVSMCs. SNHG12 silencing inhibited ox-LDL-induced proliferation and migration of hVSMCs. Moreover, SNHG12 acted as a sponge of miR-766-5p, and miR-766-5p also interacted with EIF5A. EIF5A plasmids promoted the capacities of proliferation and migration in ox-LDL-induced hVSMCs. However, shRNA-SNHG12 counteracted the facilitation of EIF5A plasmids on hVSMCs biological behaviors. CONCLUSIONS: Taken together, these findings demonstrated that silencing of SNHG12 blocks the proliferation and migration of hVSMCs via targeting the miR-766-5p/EIF5A axis.
Assuntos
MicroRNAs , RNA Longo não Codificante , Apoptose , Movimento Celular , Proliferação de Células , Humanos , Músculo Liso VascularRESUMO
Neochlorogenic acid (NCA), a natural compound found in honeysuckle, possesses prominent antiinflammatory and antitumor effects. Pingyangmycin (PYM) induces DNA damage and has been used for the treatment of oral and maxillofacial tumors. Oral care serves an important role in promoting wound healing during chemotherapy in patients with oral squamous cell carcinoma (OSCC). Therefore, the present study aimed to analyze the effects of NCA and PYM on OSCC cells and to investigate the potential underlying mechanism. Reverse transcriptionquantitative PCR and western blotting were conducted to analyze the expression levels of DNA topoisomerase II α (TOP2A) in different OSCC cell lines. TOP2Aoverexpression cells were constructed via transfection of TOP2Aoverexpression plasmids. Following NCA or PYM treatment, cell proliferation was assessed using Cell Counting Kit8 and colony formation assays, whereas cell apoptosis and the cell cycle distribution were assessed via TUNEL staining and flow cytometry, respectively. In addition, the expression levels of apoptosis and cell cyclerelated proteins were detected via western blotting. Moreover, coimmunoprecipitation (CoIP) was conducted to determine whether TOP2A interacted with CDK1. The results of the present study indicated that NCA treatment significantly enhanced the suppressive effects of PYM on OSCC cell proliferation and apoptosis. The results also indicated that PYM arrested the cell cycle in the G0/1 by regulating cyclin dependent kinase 1 (CDK1)/cyclin B1, which was enhanced by the cotreatment of NCA and PYM. In addition, NCA and PYA treatment altered the expression levels of apoptosisrelated proteins. The CoIP assay indicated that TOP2A interacted with CDK1. Moreover, TOP2A overexpression significantly reversed the effects of NCA and PYM treatment on OSCC cell proliferation and apoptosis. In addition, NCA significantly decreased PYMinduced toxicity in normal oral epithelial cells. In conclusion, the results of the present study suggested that NCA may promote the inhibitory effects of PYM in OSCC via TOP2A.
