Effect Of Epigenetic Modification Of Maspin On Extravillous Trophoblastic Function

Part OneStudy on the Expression of Maspin in Preeclamptic PlacentaObjectiveThe study detected the expression of maspin in placenta from normal and preeclamptic pregnancies.MethodsBy immunohistochemical assay and real-time PCR, the protein expression and the mRNA expression level of maspin in placenta from normal and preeclamptic pregnancies were detected.Results1. The protein of maspin was expressed in placenta of normal and preeclamptic pregnancies. The protein of maspin was strongly expressed in syncytiotrophoblast, cytotrophoblast and endothelial cell, and was mainly located in the cytoplasm.2. By contrast with normal group, the mRNA expression of maspin in preeclampsia group was significantly increased(P<0.05). Between the two groups, there was no significantly difference in maternal age, maternal height, gestational age, fetal Apgar score and fetal weight.ConclusionIn placenta, the maspin protein was mainly expressed in trophoblast. The mRNA expression level of maspin in preeclamptic placenta was significantly increased. So we speculate that abnormal expression of maspin may contribute to the pathogenesis of preeclampsia. Part Two Effect of Hypoxia on the Expression of Maspin and Biological Behavior in Extravillous TrophoblastObjectiveTo investigate the effect of hypoxia on the expression of maspin and biological behavior (include proliferation, migration and apoptosis) in extravillous trophoblast in vitro.MethodsThe study used chemical reagent (CoCl2) to establish the model of hypoxic cell. By immunocytochemistry assay, RT-PCR method, CCK8cellular proliferation assay, Transwell cellular migration assay and cellular apoptosis assay, the effect of hypoxia on the expression of maspin and biological behavior (include proliferation, migration and apoptosis) in extravillous trophoblast was detected.Results1. Maspin protein was expressed in extravillous trophoblast cell line (TEV-1), was mainly located in the cytoplasm.2. In hypoxic group, the mRNA expression of maspin in extravillous trophoblast was significantly up-regulated; the ability of proliferation and migration was obviously decreased; the apoptosis rate was significantly increased.Conclusion:The expression of maspin was increased in hypoxic trophoblast, and the aggressive behavior was obviously suppressed. The results in vitro were in accord with the phenomenon that expression of maspin in preeclamptic placenta was increased and the invasion ability of trophoblast was decreased. Therefore, we speculate that abnormal expression of maspin is related to the change of aggressive behavior of trophoblast, and play an important role in the pathogenesis of preeclampsia. Part Three Study on the Epigenetic Modification of Maspin in the Pathogenesis of PreeclampsiaObjectiveTo investigate the effect of demethylating reagent (5-Aza-dC) on the expression of maspin and biological behavior in extravillous trophoblast in vitro.MethodsBy immunocytochemistry assay, RT-PCR method, CCK8cellular proliferation assay, Transwell cellular migration assay and cellular apoptosis assay, the effect of demethylating reagent (5-Aza-dC) on the expression of maspin and biological behavior (include proliferation, migration and apoptosis) in extravillous trophoblast was detected.Results1. By contrast with PBS group, the mRNA expression of maspin in extravillous trophoblast in5-Aza-dC group was significantly increased; the ability of migration was obviously inhibited; the ability of proliferation and the rate of apoptosis had no obvious change.2. By contrast with hypoxic group, the mRNA expression of maspin in extravillous trophoblast in hypoxia+5-Aza-dC group was significantly down-regulated; the apoptosis rate in early stage was decreased; the ability of proliferation had no significant change.Conclusion:Demethylating reagent up-regulated the expression of maspin in trophoblast, and inhibited the ability of migration; demethylating reagent inhibited the effect of hypoxia on expression of maspin and apoptosis in trophoblast. The results revealed that epigenetic modification contribute to inadequate invasion of trophoblast, and provide us a potential novel direction in the study of preeclampsia.