Expression And Function Study Of Long Non-coding RNA PVT1 In Severe Preeclampsia

Objective: Preeclampsia(PE)is one of the main causes of maternal death in China,which can cause placental ischemia and hypoxia,accompanied by multiple organ damage,edema,neurological abnormalities and other complications.it is generally accepted that one of the main causes of preeclampsia is the insufficient recasting of helical arterioles,and the proliferation,migration,invasion,differentiation and apoptosis of trophoblast cells play a crucial role in the recasting of helical arterioles.Therefore,the function of trophoblast plays an important role in the occurrence and development of preeclampsia.Plasmacytoma variant translocation 1(PVT1)belongs to long non-coding RNA(lnc RNA).In recent years,a number of studies have shown that PVT1 is involved in various biological processes such as proliferation,apoptosis,migration and invasion of tumor cells,but its effect on trophoblast cells and its mechanism in severe preeclampsia have not been extensively studied and reported.Therefore,in this study,the expression levels of PVT1 in placental tissues of pregnant women with normal pregnancy and pregnant women with severe preeclampsia were detected,and the differences in expression levels between the two were analyzed,and then the effect and mechanism of PVT1 in severe preeclampsia were verified by inhibiting or overexpressing PVT1 in HTR8-S/Vneo cells.Methods:(1)Fifteen cases of pregnant women with severe preeclampsia were selected as the study subjects,and another fifteen cases of pregnant women with normal pregnancy were selected as the control group.Placental tissues were collected,and the expression gene of PVT1 in placental tissues was detected by quantitative real-time PCR(q RT-PCR).(2)After inhibition and overexpression of PVT1 in HTR8-S/Vneo cells,the proliferation,migration and apoptosis of the cells were detected by CCK8,EDU,and Western blot(WB).(3)To further explore the possible mechanism,the HTR8-S/Vneo cells were pretreated with m TOR inhibitor Rapamycin for 48 h,and then incubated with overexpression PVT1 for 48 h.Western blot was used to detect the protein expression of p-AKT,p-m TOR and apoptotic Bax,Caspase-3 and Bcl-2,and CCK8 was used to detect the proliferation of HTR8-S/Vneo cells.Conclusion:(1)The expression of PVT1 in the placental tissues of patients with severe preeclampsia was significantly lower than that of normal placental tissues.(2)After inhibition of PVT1,the proliferation and migration of HTR8-S/Vneo cells were inhibited and apoptosis was enhanced.After overexpression of PVT1,the proliferation and migration ability of HTR8-S/Vneo cells were significantly enhanced,and the apoptosis was weakened.(3)After inhibition of PVT1,the expression levels of p-AKT and p-m TOR in HTR8-S/Vneo cells decreased significantly.After overexpression of PVT1,the expression levels of p-AKT and p-m TOR in HTR8-S/Vneo cells increased significantly.(4)After the m TOR inhibitor Rapamycin(200 n M)was added to HTR8-S/Vneo cells,the overexpressed PVT1 was added and incubated for 48 h.The results showed that compared with the Over-PVT1 group,the expression levels of p-AKT and p-m TOR in the Over-PVT1+Rapamycin group were significantly decreased,and the proliferation and apoptosis of HTR8-S/Vneo cells were weakened.The above results indicate that PVT1 can affect the proliferation and apoptosis of trophoblast cells and participate in the occurrence and development of PE by regulating the AKT/m TOR signaling pathway.