rs10719 Polymorphism Located within DROSHA 3’-Untranslated Region is Responsible for Development of Primary Hypertension by Disrupting Binding with microRNA-27b
Yabing Zhang, Ai-lin Cao, Chun Dong
(Department of Cardiology, The First People’s Hospital of Jining, Jining, Shandong, China (mainland))
Med Sci Monit 2017; 23:911-918
MiR-27b is reportedly involved with many diseases (e.g., gastric cancer) by acting on different signaling pathways. In this study, we aimed at understanding the relationship between miR-27b and hypertension and its underlying molecular mechanism.
MATERIAL AND METHODS: Peripheral blood was collected from patients with hypertension, and statistical analysis was performed to study the association between rs10719 and risk of hypertension. Tissue samples were collected from patients with lung cancer, and the expression of miR-27b and DROSHA was determined using Western blot analysis and real-time PCR.
RESULTS: We first searched the miRNA database online, and identified DROSHA as a virtual target of miR-27b with the “seed sequence” located within the 3’-UTR of the target gene, and then validated DROSHA to be the direct gene via luciferase reporter assay system. We also established the negative regulatory relationship between miR-27b and DROSHA via studying the relative luciferase activity. We also conducted real-time PCR to study the mRNA and protein expression level of miR-27b among different groups. Furthermore, we conducted real-time PCR and densitometry analysis to study the mRNA and protein expression level of DROSHA among different groups of cells treated with scramble control, miR-27b mimics, DROSHA siRNA, and miR-27b inhibitors to verify the negative regulatory relationship between MiR-27b and DROSHA.
CONCLUSIONS: The presence of rs10719 disrupted the interaction between miR-27b and DROSHA, which might be the underlying mechanism of the observation that rs10719 is significantly associated with risk of primary hypertension.
Keywords: Cell Proliferation, Hypertension, MicroRNAs, Muscle, Smooth, Vascular