The Expression And Funciton Of MiR-200c In Preeclampsia Placental

Preeclampsia（PE） is a pregnancy-specific disease characterized by abnormalphysiological conversion of placental spiral arteries, which is a serious threat to the healthof both the mother and the perinatal infant. Its pathogenesis is still unknown, but it is themain reason for maternal and perinatal deaths[13]. Recent study has found that microRNA（miRNA），which is non-coding, has specific egression in human placental tissue, and itsunique function in the regulation of gene expression may be closely related to theoccurrence and development of PE [4]. Microarray study has ofund that various miRNAsare differentially expressed in preeclampsia placental tissue [5], and miR-200c is low-expressed in preeclampsia patients’ placental tissue. And related literature reported thatmiR-200family down-regulate ZEB1and ZEB2. This discovery and some recent studiesshow that the result of miR-200family directly inhibiting the expression of ZEB1andZEB2is consistent with this[6₈]. ZEB1and ZEB2are the inhibitors of E-Cad. Therefore,through the regulation of its target genes ZEB1and ZEB2，high expression of miR-200may lead to high expression of E-Cad，and then inhibit tumor metastasis by inhibition ofthe EMT process. The researchers also found that there is a negative correlation betweenmiR-200c and ZEB1and a positive correlation between miR-200c and E-Cad[9₁0]，andthen further make clear the targeting relationship between miR-200c and ZEB1and E-Cad.That ZEB1and E-Cad impact cell proliferation, invasion and apoptosis may be associatedwith the occurrence and development of PE. At present, relevant literature about miR- 200c and preeclampsia has not been reported, therefore, tfirther study of the role ofmiRNAs in the attack of PE may provide a new theoretical basis ofr the occurrence anddevelopment of PE.Objecitves:1、To validate differential expression of miR-200c in the placental tissue of normal and PEpatients;2、To make clear the targeting relationship between miR-200c and E-Cad and ZEB1.3、To imderstand the effect of miR-200c upon trophoblast cells，and make further study ofits role in preeclampsia pathogenesis.Methods:1、Using qRT-PCR to make further validation （gene chip results） that miR-200c isdifferentially expressed in preeclampsia placenta;2、In ovairan cancer and pancreatic cancer miR-200c target genes are E-Cad and ZEB1,conversion cell platform, veiryf the targeting relationship. Use qRT-PCR and Westernblot to validate target genes expression at mRNA and protein levels;3、Over express and inhibit miR-200c in HTR-8cell lines. Use cell proliferation and MTTassay to detect cell cycle，PI double staining to detect apoptosis, and Trans well to detectthe cell inifltration ability.Results:1、qRT-PCR results show that： the expression of miR-200c in preeclampsia placentaltissue was signiifcantly higher than that in normal pregnancy （/*<0.05）.2、The expression of ZEB1and E-Cad exist in both preeclampsia and normal pregnancyplacental tissue. ZEB1expression at the mRNA and protein levels was signiifcantlylower than that in normal pregnancy, while E-Cad expression at the mRNA and proteinlevels was signiifcantly higher than that in normal pregnancy. And the differences werestatistically signiifcant^°<0.05).3、In HTR-8cell lines, atfer over-expressing miR-200c，Real time PCR results show thatatfer the transfection of cells，E-Cad egression at the mRNA and protein levels was signiifcantly increased（p<0.05）, ZEB1expression at the mRNA and protein levels wassignificantly decreased（p<0.05）.4、During over expression and down-regulation of mR-200c in the HTR-8cell lines，cellproliferation and apoptosis had no signiifcant change {P>0.05). Through overexpressing mR-200c，cell inifltration capacity signiifcantly decreased（p <0.05）;through down-regulating mR-200c，cell inifltration capacity signiifcantly increased {P<0.05).Conclusion：1、 There exists imbalance expression of miRNA in preeclampsia placenta tissue, whichinvolved in the occurrence of preeclampsia and its pathophysiological process.2、As the targets of miR-200c，E-Cad and ZEB1may involve in the pathological process ofpreeclampsia by regulating cell inifltration.3、 MiR-200c involves in the occirrence and development of preeclampsia by theregulation of target genes E-Cad and ZEB1, suggests that E-Cad and ZEB1have animportant role in the pathogenesis of preeclampsia, and can provide a new theoreticalbasis for the study of the pathogenesis of preeclampsia.