miR-191 inhibits retinal vascular endothelial cell proliferation and angiogenesis
10.3760/cma.j.cn511434-20210406-00176
- VernacularTitle:miR-191抑制视网膜血管内皮细胞增生和血管形成
- Author:
Qi ZHAO
1
;
Weiting AN
;
Boshi LIU
;
Manhong XU
;
Lijie DONG
;
Xiaorong LI
;
Jindong HAN
Author Information
1. 天津医科大学眼科医院、眼视光学院、眼科研究所 国家眼耳鼻喉疾病临床医学研究中心天津市分中心 天津市视网膜功能与疾病重点实验室，天津 300384
- Keywords:
MicroRNAs;
Endothelium, vascular;
Transfection;
Hyperplasia
- From:
Chinese Journal of Ocular Fundus Diseases
2022;38(1):49-55
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To observe the inhibitory effect of lentivirus (LV)-mediated miR-191 on the proliferation and angiogenesis of human retinal vascular endothelial cells (hREC) cultured in vitro.Methods:The hREC cell lines were cultured in vitro and divided into control group, hypoxia group, LV-empty vector (LV-vector) group, and LV-miR-191 (LV-191) group. The LV-vector group and LV-191 group were transferred to the corresponding lentiviral vector respectively. Flow cytometry was used to detect cell transfection efficiency. Cell Counting Kit-8 (CCK-8) test was used to detect cell proliferation ability. Scarification test and invasion chamber (Transwell) test were used to detect cell migration ability. Matrigel test was used to detect cell lumen formation ability. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the relative expression of miR-191 and relative mRNA expression of its downstream target genes p21, vascular endothelial growth factor (VEGF), cell division protein kinase (CDK) 6, cyclin-D1 (Cyclin D1). Independent sample t test was used for pairwise comparison. Results:The results of flow cytometry showed that the transfection efficiency of cells in the control group and the LV-191 group were 0.615% and 99.400%, respectively. The results of CCK-8, scarification, Transwell and Matrigel test showed that, compared with the control group, the number of cell proliferation ( t=6.130, 4.606), the cell mobility ( t=4.910, 6.702), the number of stained cells on the microporous membrane ( t=7.244, 6.724) and the lumen formation ability cells ( t=8.345, 9.859) were significantly increased in the hypoxia group and the LV-vector group ( P<0.01), while the LV-191 group showed completely opposite performance ( t=14.710, 6.245, 5.333, 5.892; P≤0.01). The qPCR test results showed that, compared with the control group and the LV-vector group, the relative expression of miR-191 mRNA in the cells of the LV-191 group was significantly up-regulated ( t=44.110, 42.680), the relative expression of Cyclin D1 mRNA ( t=29.940, 14.010) and CDK6 mRNA ( t=15.200, 7.645) decreased significantly, and the difference were statistically significant ( P<0.01); the relative expression of p21 mRNA increased, however, the difference was not statistically significant ( t=2.013, 2.755; P>0.05). There was no significant difference in the relative expression of VEGF mRNA in the 4 groups of cells ( F=0.966, P>0.05). Conclusions:LV-191 can inhibit the proliferation, migration and tubing of hREC by up-regulating p21 and down-regulating CDK6 and Cyclin D1.
