Jin Xu1, Yanqing Xia1, Helong Zhang1, Haibin Guo1, Ke Feng1, Cuilian Zhang2. 1. Reproductive Medicine Center, Henan Provincial People's Hospital, Zhengzhou University, Zhengzhou 450003, PR China. 2. Reproductive Medicine Center, Henan Provincial People's Hospital, Zhengzhou University, Zhengzhou 450003, PR China. Electronic address: positi.Mattns0azyl@gmail.com.
Abstract
BACKGROUND: Preeclampsia (PE), characterized by hypertension and proteinuria, is a leading cause of perinatal and maternal mortality. Considering that mutation of H19 gene is closely associated with PE, we aimed to explore the functional role of long non-coding RNA H19 (lncRNA-H19) in trophoblast cells. METHODS: Expression of lncRNA-H19 in placenta tissues from patients with PE and healthy pregnant women after delivery was determined by quantitative reverse transcription PCR. Then, lncRNA-H19 was abnormally expressed in JEG-3 and HTR-8 cells by stable cell transfection. Cell viability and invasion were assessed by using CCK-8 and Matrigel-coated Millicell system, respectively. Expression of key proteins associated with invasion and autophagy as well as key kinases in the phosphatidylinositol-3-kinase (PI3K)/AKT/mechanistic target of rapamycin (mTOR) pathways were measured by Western blot analysis. Number of GFP-labeled autophagosomes was counted under a confocal microscope. RESULTS: Level of lncRNA-H19 in the placenta tissues from PE patients was higher than that from healthy controls. LncRNA-H19 overexpression reduced cell viability but increased invasion of JEG-3 and HTR-8 cells. LncRNA-H19 silence showed the opposite effects. In addition, lncRNA-H19 overexpression promoted autophagy in trophoblast cells. Furthermore, phosphorylated levels of key kinases in the PI3K/AKT/mTOR pathways were enhanced by lncRNA-H19 overexpression while were reduced by lncRNA-H19 silence. CONCLUSION: LncRNA-H19, which was up-regulated in PE, reduced cell viability but promoted invasion and autophagy in trophoblast cells, along with activation of the PI3K/AKT/mTOR pathways. Our study provides a theoretical basis for pathogenesis of PE, aiding to identification of novel therapeutic strategies for PE.
BACKGROUND: Preeclampsia (PE), characterized by hypertension and proteinuria, is a leading cause of perinatal and maternal mortality. Considering that mutation of H19 gene is closely associated with PE, we aimed to explore the functional role of long non-coding RNA H19 (lncRNA-H19) in trophoblast cells. METHODS: Expression of lncRNA-H19 in placenta tissues from patients with PE and healthy pregnant women after delivery was determined by quantitative reverse transcription PCR. Then, lncRNA-H19 was abnormally expressed in JEG-3 and HTR-8 cells by stable cell transfection. Cell viability and invasion were assessed by using CCK-8 and Matrigel-coated Millicell system, respectively. Expression of key proteins associated with invasion and autophagy as well as key kinases in the phosphatidylinositol-3-kinase (PI3K)/AKT/mechanistic target of rapamycin (mTOR) pathways were measured by Western blot analysis. Number of GFP-labeled autophagosomes was counted under a confocal microscope. RESULTS: Level of lncRNA-H19 in the placenta tissues from PE patients was higher than that from healthy controls. LncRNA-H19 overexpression reduced cell viability but increased invasion of JEG-3 and HTR-8 cells. LncRNA-H19 silence showed the opposite effects. In addition, lncRNA-H19 overexpression promoted autophagy in trophoblast cells. Furthermore, phosphorylated levels of key kinases in the PI3K/AKT/mTOR pathways were enhanced by lncRNA-H19 overexpression while were reduced by lncRNA-H19 silence. CONCLUSION: LncRNA-H19, which was up-regulated in PE, reduced cell viability but promoted invasion and autophagy in trophoblast cells, along with activation of the PI3K/AKT/mTOR pathways. Our study provides a theoretical basis for pathogenesis of PE, aiding to identification of novel therapeutic strategies for PE.