Long intergenic ncRNA 00473 improves the invasion of trophoblastic cells via miR-16-5p.

Preeclampsia (PE) is a common disease among pregnant women and is characterized by high blood pressure, edemas, proteinuria, etc. However, the underlying mechanism of PE is still not clear. Our results may provide a new understanding of the pathogenesis of PE and a therapeutical target for the treatment of the disease. Levels of long intergenic ncRNA 00473 (LINC00473), miR-16-5p, MMP2, MMP9, Bcl-2, Bax, and C caspase-3 in placental tissues or human trophoblastic cells were assessed. HTR8/SVneo and JEG-3 cells were transfected with LINC00473, miR-16-5p mimic, LINC00473 siRNA, or miR-16-5p inhibitor alone, or co-transfected with LINC00473 and miR-16-5p mimic or LINC00473 siRNA and miR-16-5p inhibitor. Viability, apoptosis, migration and invasion of cells were assessed by Cell Counting Kit-8, flow cytometry, wound healing assay and Transwell assay, respectively. The target gene of LINC00473 was analyzed using Starbase and dual-luciferase reporter assay. LINC00473 level was down-regulated in placental tissues of PE patients. LINC00473 overexpression increased cell viability, migration, invasion, and MMP2, MMP9 and Bcl-2 levels, yet decreased the apoptosis rates and Bax and C caspase-3 levels in cells; however, LINC00473 silencing had the opposite effect. LINC00473 targeted miR-16-5p and miR-16-5p level was negatively related to LINC00473 level. MiR-16-5p mimic reversed the promoting effect of LINC00473 overexpression on the invasion of HTR8/SVneo and JEG-3 cells, while miR-16-5p inhibitor reversed the inhibitory effect of LINC00473 silencing on the invasion of these cells. In conclusion, LINC00473 improved the invasion of human trophoblastic cells via miR-16-5p.

Effect of SHBG gene on the apoptosis of human trophoblastic cells / 局解手术学杂志

Objective To investigate the effects of sex hormone-binding globulin (SHBG)gene in the apoptosis of human trophoblastic cells.Methods The siRNA specific-targeting SHBG gene was transfected into human trophoblastic cells and they were divided into six groups:trophoblasts without transfection in normal control groups(group Ⅰ);transfect liposome in blank control groups(group Ⅱ);transfect nonspecific siRNA in negative control groups(group Ⅲ);transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively in trans-fection group(group Ⅳ,Ⅴ,Ⅵ).Hoechst 33258 dying method was used to detect cell apoptosis.SHBG and Caspase-3 mRNA profiling and the level of SHBG and caspase-3 protein were detected by real-time PCR and Western blot.Results There was no statistical significant difference in the gene expression and protein level of SHBG and caspase-3 in group Ⅰ,Ⅱ and Ⅲ (P >0.05).In Ⅳ,Ⅴ and Ⅵ group,there was no statistical significant difference in the expression level of SHBG and caspase 3 (P >0.05).Compared with group Ⅰ,Ⅱ and Ⅲ,the a-mount of SHBG gene expression decreased obviously,the caspase-3 mRNA and protein level increased obviously and the trophoblast cell ap-optosis increased markedly (P <0.05).Conclusion Through siRNA interference technology can reduce SHBG gene expression in human trophoblastic cells,and it can lead to excessive apoptosis of human trophoblasts cells.

Exposure to ethanol and nicotine induces stress responses in human placental BeWo cells.

Human placental trophoblastic cancer BeWo cells can be used as a model of placental trophoblasts. We found that combined exposure to relevant exposure concentrations of ethanol (2‰) and nicotine (15 µM) induces an increase in the amount of reactive oxygen species (ROS). Neither ethanol or nicotine alone, nor their combination affected cell viability. However, nicotine decreased cell proliferation, both alone and combined with ethanol. Nicotine increased the expression of the endoplasmic reticulum (ER)-stress related protein GRP78/BiP, but not another marker of ER-stress, IRE1α. We also studied the effects of nicotine and/or ethanol on phosphorylation and expression of three mitogen-activated protein kinases (MAPKs), i.e. JNK, p38 and ERK1/2. Nicotine decreased the phosphorylation of JNK and also had similar effect on total amount of this protein. Phosphorylation and expression of p38 were increased 1.7- and 1.6-fold, respectively, by nicotine alone, and 1.9- and 2.1-fold by the combined treatment. Some increase (1.8-fold) was also seen in the phosphorylation of ERK2 at 48 h, in cells exposed to both ethanol and nicotine. This study shows that ethanol and nicotine, which harm the development of fetus may induce both oxidative and ER stress responses in human placental trophoblastic cells, implicating these mechanisms in their fetotoxic effects.