Study Of MiR-326 Targeting PAX8 To Inhibit The Proliferation And Invasion Of Trophoblast Cells

Objective:To explore the role of mir-326 in regulating trophoblast proliferation and invasion in the pathogenesis of preeclampsia,and to provide a new theoretical basis for the diagnosis and treatment of preeclampsia.Methods:1.Specimen:(1)The placental tissues of normal pregnant women(n=6)and pre-eclampsia pregnant women(n=6)were collected.(2)Nourish cells HTR-8/SVneo and JE-3.2.Experimental method:(1)The expression level of miR-326 in placenta was detected by q RT-PCR.(2)Transwell,CCK-8 and clone formation experiments were used to detect the invasion and proliferation of transfected trophoblasts.(3)Online database Target Scan predicts the possible target genes of miR-326.(4)The expression levels of miRNA and protein of related molecules were detected by q RT-PCR and Western blot.(5)Establish overexpression and knock-down cell lines of target protein.(6)Using miR-326 analogue and PAX8 overexpression lentivirus to treat and transfect trophoblast at the same time,and detect the invasion and proliferation of trophoblast.Results:1.The expression level of miR-326 in placenta of preeclampsia patients was significantly higher than that of normal pregnant women.2.MiR-326 analogues can overexpress MIR-326 in trophoblast HTR-8/SVneo,and significantly inhibit the proliferation and invasion of trophoblast.MiR-326 inhibitor can inhibit the expression of miR-326 in trophoblast,and significantly enhance the proliferation and invasion of JEG-3.3.Online databases(Target Scan,miRDB and micro RNA.org)predicted that the downstream target gene of miR-326 was PAX8.4.The transfection efficiency of PAX8 overexpression lentivirus and knock-down vector is high.5.The proliferation and invasion ability of PAX8-overexpressing trophoblast JEG-3increased significantly,while the proliferation and invasion ability of PAX8-knocked down trophoblast HTR-8/SVneo decreased significantly.6.The trophoblast HTR-8/SVneo treated with miR-326 analogue was transfected into PAX8 overexpression lentivirus.The result showed that the expression of miR-326 was increased,and its inhibition on trophoblast proliferation and invasion could be reversed by the overexpression of PAX8.Conclusions:1.MiR-326 can significantly inhibit the proliferation and invasion of trophoblasts.2.MiR-326 can inhibit the proliferation and invasion of trophoblasts by targeting regulation of PAX8 in trophoblasts.