Investigation Of The Regulation Of LncRNA TINCR In Placental Tissue Of Patients With Preeclampsia And The Mechanisms Underlying Its Effect On Trophoblasts

Objective: To investigate the expression and clinical significance of terminal differentiation-induced non-coding RNA(TINCR),Wnt5 a protein and ?-catenin protein in placental tissues of patients between severe preeclampsia(SPE)and control group.And to explore its mechanism of research on the biological behavior of trophoblasts.Methods: 1.Thirty pregnant patients with severe preeclampsia(SPE)at caesarean delivery in our hospital from April 2016 to April 2018 were selected as severe preeclampsia group.Thirty pregnancy pregnant women were in the control group.qRT-PCR was used to detect the relative expression of TINCR In the placenta.The expression and localization of Wnt5 a protein and ?-catenin protein in placenta were detected by Western blot and immunohistochemistry.2.HTR-8/SVneo trophoblasts were cultured in vitro,and an overexpression plasmid was constructed.The plasmids transfected with TINCR and TINCR siRNA were transfected into HTR-8/SVneo cells using lipo3000 and/or P3000.The transfection efficiency was examined by qRT-PCR.The proliferationability of HTR-8/SVneo was detected by MTT assay,the migration ability of HTR-8/SVneo was detected by scratch test,and the infiltration ability of HTR-8/SVneo was detected by Transwell assay.Western blot was used to detect the expression levels of Wnt5 a protein and ?-catenin protein in the downstream of TINCR,and further study the effect of TINCR on the biological function of trophoblasts through Wnt5 a protein and ?-catenin protein.Results:1.we used qRT-PCR to detect the placental tissue of the preeclampsia group and the control group and found that TINCR was highly expressed in the preeclamptic placenta tissue.We used western blot to show that the expression of Wnt5 a protein and ?-catenin proteinwasdecreased in the preeclamptic placenta tissue,compared with the control group.We used immunohistochemistry to further determine the expression sites of Wnt5 a and ?-catenin in placental tissues,and found that Wnt5 a and ?-catenin proteins can be expressed in cytoplasm of syncytiotrophoblasts and cytotrophoblasts.At the same time,it was found that TINCR is very likely to participate in the biological behavior of trophoblasts,which may cause the onset of preeclampsia.2.Inorder to study the potential biological behavior of TINCR on HTR-8/SVneo,we established Si-TINCR and PX856-TINCR transfection systems,and transferred TINCR expression into HTR-8/SVneo cells from Si-TINCR.The level was significantly down-regulated(P < 0.001),and the expression of TINCR was significantly up-regulated in HTR-8/SVneo cells transfected with PX856-TINCR plasmid(P <0.01),compared with the negative control group and the blank control group.The proliferation ability of HTR-8/SVneo cells was detected by MTT assay.In the knockdown experiment,we found that the Si-TINCR group began to increase after 48h(P <0.01).In the overexpression experiment,we found that the proliferation of PX856-TINCR group decreased.(P <0.01).The migration function of HTR-8 /SVneo cells was detected by cell scratch test.In the knockdown experiment,we found that the mobility of the si-TINCR group increased(P < 0.001).In the overexpression experiment,we found that the mobility of the PX856-TINCR group decreased.(P <0.001).The invasion ability of HTR-8/SVneo cells was detected by Transwell assay.In the knockdown experiment,we found that the number of invading cells in the si-TINCR group increased(P< 0.001).In the overexpression experiment,we found that the number of invading cells in the PX856-TINCR group decreased(P <0.01).Conclusion: 1.In placental tissue of preeclampsia,high expression of TINCR is associated with low expression of Wnt5 a protein and ?-catenin protein,which may be involved in the pathogenesis of preeclampsia.2.TINCR participates in the biological behavior of HTR-8/SVneo cells.3.TINCR can inhibit the proliferation,migration and infiltration of HTR-8/SVneo cells.4.TINCR may influence the proliferation,migration and infiltration of trophoblast cells through the combination of classical Wnt signaling pathway or non-canonical Wnt signaling pathway or classical and non-canonical Wnt signaling pathways,and participate in the pathogenesis of preeclampsia.