Ge Lihao, Wang Yuyan, Cao Yang, Li Gang, Sun Rui, Teng Peng, Wang Yansong, Bi Yunlong, Guo Zhanpeng, Yuan Yajiang, Yu Deshui
Department of Orthopedics, The First Affiliated Hospital, Jinzhou Medical University, Jinzhou, 121001, China.
Department of Gynaecology and Obstetrics, The First Affiliated Hospital, Jinzhou Medical University, Jinzhou, 121001, China.
Biotechnol Lett. 2018 Dec;40(11-12):1477-1486. doi: 10.1007/s10529-018-2604-6. Epub 2018 Aug 25.
MicroRNA-429(miR-429) plays an important role in mesenchymal stem cells. Hypoxia-inducible factor 1α (HIF-1α) is a nuclear transcription factor that regulates the proliferation, apoptosis and tolerance to hypoxia of mesenchymal stem cells. HIF-1α is also a target gene of miR-429. We investigated whether miR-429 plays a role in hypoxia tolerance with HIF-1α in human amniotic mesenchymal stem cells (hAMSCs). The expression of miR-429 was increased by hypoxia in hAMSCs. miR-429 expression resulted in decreased HIF-1α protein level, but little effect on HIF-1α mRNA. While overexpression of HIF-1α increased the survival rate and exhibited anti-apoptosis effects in hAMSCs under hypoxia, co-expression of miR-429 reduced survival and increased apoptosis. However, miR-429 silencing with HIF-1α overexpression stimulated cell survival and reduced apoptosis. Co-expression of HIF-1α and miR-429 reduced VEGF and Bcl-2 proteins and increased Bax and C-Caspase-3 levels in hAMSCs under hypoxia compared with cells expressing only HIF-1α; cells with HIF-1α overexpression and miR-429 silencing showed the opposite effects. These results indicate that HIF-1α and angomiR-429 reciprocally antagonized each other, while HIF-1α and antagomiR-429 interacted with each other to regulate survival and apoptosis in hAMSCs under hypoxia. miR-429 increased VEGF and Bcl-2 protein levels and decreased Bax and cleaved Caspase-3 protein levels by promoting the synthesis of HIF-1α. These results indicate that miR-429 negatively regulates the survival and anti-apoptosis ability of hAMSCs by mediating HIF-1α expression and improves the ability of hAMSCs to tolerate hypoxia.
微小RNA-429（miR-429）在间充质干细胞中发挥重要作用。缺氧诱导因子1α（HIF-1α）是一种核转录因子，可调节间充质干细胞的增殖、凋亡及缺氧耐受性。HIF-1α也是miR-429的靶基因。我们研究了miR-429在人羊膜间充质干细胞（hAMSCs）中是否通过HIF-1α发挥缺氧耐受作用。缺氧可使hAMSCs中miR-429的表达增加。miR-429表达导致HIF-1α蛋白水平降低，但对HIF-1α mRNA影响较小。虽然HIF-1α过表达可提高hAMSCs在缺氧条件下的存活率并表现出抗凋亡作用，但miR-429共表达则降低了存活率并增加了凋亡。然而，miR-429沉默与HIF-1α过表达共同作用可促进细胞存活并减少凋亡。与仅表达HIF-1α的细胞相比，缺氧条件下hAMSCs中HIF-1α与miR-429共表达可降低VEGF和Bcl-2蛋白水平，并增加Bax和C-Caspase-3水平；HIF-1α过表达且miR-429沉默的细胞则表现出相反的效果。这些结果表明，HIF-1α与抗miR-429相互拮抗，而HIF-1α与抗miR-429相互作用以调节hAMSCs在缺氧条件下的存活和凋亡。miR-429通过促进HIF-1α的合成增加VEGF和Bcl-2蛋白水平，并降低Bax和裂解的Caspase-3蛋白水平。这些结果表明，miR-429通过介导HIF-1α表达负向调节hAMSCs的存活和抗凋亡能力，并提高hAMSCs的缺氧耐受能力。
