RESUMEN
BACKGROUND: Pre-eclampsia (PE) is regarded as the leading cause of maternal and neonatal morbidity and mortality. Nevertheless, the potential mechanism for the regulation of trophoblast behaviors and the pathogenesis of PE remain largely elusive. Recently, accumulating evidence emphasized that aberrant expression of long non-coding RNAs (lncRNAs) functions as imperative regulators in human diseases, including PE. Thus, identifying PE-related specific lncRNAs to uncover the underlying molecular mechanism is of much significance. However, the functional roles and underlying mechanisms of lncRNAs in PE progression remain unclear. METHOD: Placenta tissues obtained from patients with PE and healthy pregnant women were performed to measure TUG1 expression by qRT-PCR analysis. Transient transfections were conducted to alter TUG1 expression. Cell Counting Kit-8 (CCK-8) and flow cytometry assays were carried out to assess cell proliferation and apoptosis, respectively. Transwell and tube formation assays were performed to measure the capacity of cell invasion and angiogenesis. Moreover, the luciferase reporter assay was subjected to verify the binding relationship between TUG1 and miR-29b. Western blot analysis was performed to detect the expression of key proteins in the PI3K/AKT and ERK pathway. RESULTS: Here, we identified a lncRNA, TUG1, which was notably decreased in placental samples of PE patients. Functional experiments of loss- or gain-of-function assays also verified that ectopic expression of TUG1 promoted cell proliferation, invasion, and angiogenesis, but negatively regulated cell apoptosis, whereas TUG1 inhibition presented the opposite effects. Furthermore, mechanistic researches revealed that TUG1 could act as a molecular sponge for miR-29b, thus regulating MCL1, VEGFA, and MMP2 to modulate PE development. CONCLUSIONS: Taken together, our findings demonstrated that TUG1 exerts as a critical role in PE progression, which might furnish a novel therapeutic marker for PE treatment.
Asunto(s)
Proliferación Celular/genética , MicroARNs/genética , Preeclampsia/genética , ARN Largo no Codificante/genética , Apoptosis/genética , Línea Celular , Movimiento Celular/genética , Femenino , Humanos , Metaloproteinasa 2 de la Matriz/genética , Proteína 1 de la Secuencia de Leucemia de Células Mieloides/genética , Neovascularización Patológica/genética , Neovascularización Patológica/patología , Placenta/metabolismo , Placenta/patología , Preeclampsia/patología , Embarazo , Trofoblastos/metabolismo , Trofoblastos/patología , Factor A de Crecimiento Endotelial Vascular/genéticaRESUMEN
OBJECTIVE: To study the effects of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotide (ODN) on the preeclamptic umbilical serum induced expression of precollagen I, III mRNA and tumor necrosis factor-alpha (TNF-alpha) in cultured human umbilical artery smooth muscle cells (HUASMC). METHODS: Primary cultured HUASMC of normal pregnancy were divided into four groups: group A (HUASMC were incubated with umbilical serum of normal pregnancy); group B (HUASMC were incubated with umbilical serum of preeclampsia); group C (HUASMC were transfected with NF-kappaB cis decoy ODN 48 h before incubation with umbilical serum of preeclampsia); group D (HUASMC were transfected with NF-kappaB scramble ODN 24 h before incubation with umbilical serum of preeclampsia). NF-kappaB cis decoy ODN and NF-kappaB scramble ODN were transfected with cationic lipofectamine to the latter two groups, respectively. The proliferation of human umbilical artery smooth muscle cells was evaluated by methyl thiazolyl tetrazolium and the apoptosis was analyzed by flow cytometry. The expression levels of precollagen I, III mRNA were detected by RT-PCR, the expression levels of TNF-alpha were detected by western blot. RESULTS: (1) The proliferation of group B (0.19 +/- 0.02) and group D (0.18 +/- 0.03) was significantly increased as compared with those of group A (0.11 +/- 0.02) and group C (0.14 +/- 0.02) (P < 0.05). (2) The apoptosis rates of group B [(7.8 +/- 1.3)%], group C [(10.1 +/- 1.2)%] and group D [(8.1 +/- 1.3)%] were significantly reduced as compared with that of group A [(14.3 +/- 1.2)%] (P < 0.05), and there was a significant difference between groups B and C (P < 0.05). (3) The expression levels of precollagen I mRNA of group B (0.31 +/- 0.04), group C (0.23 +/- 0.04) and group D (0.30 +/- 0.03) were significantly increased as compared with that of group A (0.16 +/- 0.02) (P < 0.05), and there was a significant difference between groups B and C (P < 0.05). (4) There were no significant differences among the four groups in the expression level of precollagen III mRNA (P > 0.05). (5) The expression of TNF-alpha of group B (0.74 +/- 0.11), group C (0.36 +/- 0.09) and group D (0.79 +/- 0.12) were significantly higher than that of group A (0.15 +/- 0.03) (P < 0.05), and the expression of TNF-alpha of groups B and D were significantly higher than that of group C (P < 0.05), while there was no significant difference between groups B and D (P > 0.05). CONCLUSIONS: NF-kappaB cis decoy ODN could down-regulate the proliferation, as well as the expression levels of precollagen and TNF-alpha of HUASMC induced by umbilical serum of preeclampsia. NF-kappaB may play an important role in the pathogenesis of placental artery abnormalities in preeclampsia.