Wang Jian, Zhao Xiangzhai, Guo Zhaojun, Ma Xiaolin, Song Yueqing, Guo Ying
Department of Obstetric and Gynecology, The Third Hospital of Hebei Medical University, 139 Ziqiang Rd., Shijiazhuang, Hebei, 050051, People's Republic of China.
Arch Gynecol Obstet. 2017 Jun;295(6):1469-1475. doi: 10.1007/s00404-017-4365-1. Epub 2017 Apr 26.
To investigate the function and mechanism of lnc NEAT1 in regulating the growth, migration and invasion of endometrial carcinoma (EC) cells.
NEAT1 and miR-214-3p levels were measured by qRT-PCR. The protein levels of HMGA1, β-catenin, c-myc and MMP9 were evaluated by Western blot. The effects of NEAT1, HMGA1, miR-214-3p on the viability, migration and invasion of HEC-1A cells were accessed by WST-1 assay and transwell migration/invasion assay. The effect of miR-214-3p on Wnt signaling activity was tested by luciferase reporter assay.
NEAT1, HMGA1 and β-catenin were significantly upregulated in EC tissues, and miR-214-3p was significantly downregulated. NEAT1 promoted the growth, migration and invasion of HEC-1A cells, and mRNA level of Wnt/β-catenin downstream genes c-myc and MMP9. In addition, HMGA1 upregualted the protein and mRNA levels of Wnt/β-catenin downstream genes c-myc and MMP9, and could improve cell viability, and increase numbers of migration and invasion of HEC-1A cells. miR-214-3p overexpression inhibited the proliferation, migration and invasion of HEC-1A cells, while NEAT1 overexpression reversed these effects. miR-214-3p overexpression inhibited the activity of Wnt/β-catenin pathway, while NEAT1 overexpression reversed this effect. Then, si-HMGA1 reduced the activity of Wnt/β-catenin pathway. Moreover, we found NEAT1 and HMGA1 bound to miR-214-3p by luciferase reporter assay, and NEAT1 and HMGA1 expression were negatively correlated with miR-214-3p.
NEAT1 regulates HMGA1 via miR-214-3p to regulate Wnt/β-catenin pathway, thus promotes the growth, migration and invasion of HEC-1A cells.
探讨长链非编码RNA NEAT1在调控子宫内膜癌（EC）细胞生长、迁移和侵袭中的作用及机制。
采用qRT-PCR检测NEAT1和miR-214-3p水平。通过蛋白质印迹法评估HMGA1、β-连环蛋白、c-myc和MMP9的蛋白水平。采用WST-1法和Transwell迁移/侵袭实验检测NEAT1、HMGA1、miR-214-3p对HEC-1A细胞活力、迁移和侵袭的影响。通过荧光素酶报告基因实验检测miR-214-3p对Wnt信号活性的影响。
NEAT1、HMGA1和β-连环蛋白在EC组织中显著上调，而miR-214-3p显著下调。NEAT1促进HEC-1A细胞的生长、迁移和侵袭，并上调Wnt/β-连环蛋白下游基因c-myc和MMP9的mRNA水平。此外，HMGA1上调Wnt/β-连环蛋白下游基因c-myc和MMP9的蛋白及mRNA水平，并可提高细胞活力，增加HEC-1A细胞的迁移和侵袭数量。miR-214-3p过表达抑制HEC-1A细胞的增殖、迁移和侵袭，而NEAT1过表达可逆转这些作用。miR-2过表达抑制Wnt/β-连环蛋白通路的活性，而NEAT1过表达可逆转此作用。然后，si-HMGA1降低了Wnt/β-连环蛋白通路的活性。此外，通过荧光素酶报告基因实验发现NEAT1和HMGA1与miR-214-3p结合，且NEAT1和HMGA1的表达与miR-214-3p呈负相关。
NEAT1通过miR-214-3p调控HMGA1以调节Wnt/β-连环蛋白通路，从而促进HEC-1A细胞的生长、迁移和侵袭。
