Effects of miR-99a-mTOR in the regulation of human umbilical vein endothelia cell proliferation under hypoxia
10.3760/cma.j.issn.1009-6906.2017.02.012
- VernacularTitle:miR-99a-mTOR在低氧调控人脐静脉内皮细胞增殖中的作用研究
- Author:
Li ZHANG
1
;
Hao ZHANG
;
Zhaogang DONG
;
Zhaoquan XING
Author Information
1. 中国矿业大学体育系, 北京,100083
- Publication Type:Journal Article
- Keywords:
Hypoxia;
MiR-99a;
mTOR;
Proliferation
- From:
Chinese journal of nautical medicine and hyperbaric medicine
2017;24(2):130-134
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of hypoxia on the proliferation of human umbilical vein endothelia cell (HUVEC) and its underlying mechanism.Methods HUVEC were incubated at different oxygen concentrations (1％,5％,21％) and at 5％ CO2 at a temperature of 37 ℃ for 24 hours.Proliferation rate was detected by WST-1.The target gene of miR-99a was predicted by bioinformatics and verified by double luciferase reporter assay.MiR-99a mimics,miR-99a siRNA,mTOR siRNA,or mTOR cDNA were transfected in HUVEC by lipofectamine 2000,and then incubated in hypoxia.Mir-99a and mTOR were respectively detected by qRT-PCR and RT-PCR or Western blot.Results The proliferation of HUVEC was obviously decreased under hypoxia (1％ O2) (33.3％ ± 6.5％),as compared with normoxia(21％ 02) (P =0.004).Hypoxia could up-regulate the expression level of miR-99a (1.94 ± 0.24,P =0.001),which further inhibited the proliferation of HUVEC.Bioinformatics and double luciferase reporter assay indicated that mTOR was the target gene of miR-99a.Under hypoxia,the gene and protein level of mTOR were down-regulated by miR-99a.Further research demonstrated that the proliferation of HUVEC was inhibited under hypoxia by miR-99a-mTOR pathway.Conclusions Hypobaric treatment could significantly inhibit the proliferation of HUVEC,which might be associated with miR-99a-mTOR pathway.
