Function and mechanism of microRNA-210 in acute cerebral infarction

Abstract

Acute cerebral infarction (ACI) is a common cerebrovascular disease. Previous studies have indicated that microRNAs (miRs) are aberrantly expressed in patients with ACI. However, the functions of miRs in the pathogenesis of ACI still require further investigation. The aim of the present study was to investigate the function of miR-210 in ACI and its associated mechanism. The expression of miR-210 in the serum of 40 patients with ACI and 40 normal controls was examined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Then, human umbilical vein endothelial cells (HUVECs) were treated with serum from patients with ACI or healthy volunteers, and a CCK-8 assay was performed to examine cell proliferation. Next, cells were stained with PI/Annexin V, and the apoptosis rate was examined using flow cytometry. Furthermore, cells were harvested and lysed, and RT-qPCR and western blotting assays were performed to compare the expression of vascular endothelial growth factor (VEGF), Notch1 and Hes1 in different groups. It was observed that the expression of miR-210 was significantly increased in the serum of patients with ACI compared with normal controls (P<0.01), and receiver operating characteristic curve analysis indicated that the area under the curve for miR-210 was 0.799 (95% confidence interval, 0.700-0.899), the optimum cut-off point was 1.397, and the sensitivity and specificity at the cut-off point were 62.5 and 87.5%, respectively. Furthermore, serum from patients with ACI induced a significant increase in proliferation (P<0.05 at 48 h, P<0.01 at 72 h) and a significant decrease in the apoptosis rate of HUVECs (P<0.01). In addition, serum from patients with ACI significantly increased the expression of VEGF, Notch1 and Hes1 at the mRNA and protein level (all P<0.01 with the exception of Notch1 mRNA expression, P>0.05). In conclusion, these results demonstrate that miR-210 is upregulated in the serum of patients with ACI, and miR-210 may be involved in the pathogenesis of ACI through regulating the proliferation and apoptosis of endothelial cells.

Keywords: acute cerebral infarction; apoptosis; human umbilical vein endothelial cells; micoRNA-210; proliferation.

Figure 1.

Expression of miR-210 in ACI. (A) Relative expression of miR-210 in the serum of patients with ACI and healthy controls; (B) Receiver operating characteristic curve analysis of miR-210. ***P<0.001 vs. healthy controls. miR, microRNA; ACI, acute cerebral infarction.

Figure 2.

Expression of miR-210 in human umbilical vein endothelial cells treated with different mediums. **P<0.01 vs. FBS. FBS, fetal bovine serum; miR, microRNA.

Figure 3.

Proliferation rate of cells in different groups. *P<0.05, **P<0.01 vs. control. Control, untransfected cells; NC, cells transfected with negative control miR-210 mimics; miR-210, cells transfected with miR-210 mimics; miR, microRNA.

Figure 4.

Apoptosis of cells in different groups: (A) Control, (B) NC and (C) miR-210. (D) Apoptosis rate was determined in each group. **P<0.01 vs. control. Control, untransfected cells; NC, cells transfected with negative control miR-210 mimics; miR-210, cells transfected with miR-210 mimics; miR, microRNA.

Figure 5.

Relative mRNA expression of (A) Hes1, (B) Notch1 and (C) VEGF in human umbilical vein endothelial cells in different groups. **P<0.01 vs. control. Control, untransfected cells; NC, cells transfected with negative control miR-210 mimics; miR-210, cells transfected with miR-210 mimics; miR, microRNA; VEGF, vascular endothelial growth factor.

Figure 6.

(A) Protein expression of Hes1, Notch1 and VEGF in human umbilical vein endothelial cells in different groups. (B) Relative protein expression levels were semi-quantified. **P<0.01 vs. control. Control, untransfected cells; NC, cells transfected with negative control miR-210 mimics; miR-210, cells transfected with miR-210 mimics; miR, microRNA; VEGF, vascular endothelial growth factor.