Abstract
Preterm labor (PTL) is one of the obstetric complications, and is known to be associated with abnormal maternal inflammatory response and intrauterine inflammation and/or infection. However, the expression of specific miRNAs associated with PTL is not clear. In this study, we performed combination analysis of miRNA array and gene array, and then selected one miRNA (miR-373-3p) and its putative target genes (CD44 and RDX) that exhibited large expression differences in term and PTL placentas with or without inflammation. Using qRT-PCR and luciferase assays, we confirmed that miR-373-3p directly targeted CD44 and RDX. Overexpression of miR-373-3p reduced the migration and invasion of trophoblast cells, while inhibition of miR-373-3p restored the migration and invasion abilities of trophoblast cells. Finally, we validated the expression of miR-373-3p and its target genes in clinical patients’ blood. miR-373-3p was increased in PTL patients’ blood, and was the most expressed in PTL patients’ blood with inflammation. In addition, by targeting the miR-373-3p, CD44 and RDX was decreased in PTL patients’ blood, and their expression were the lowest in PTL patients’ blood with inflammation. Taken together, these findings suggest that miR-373-3p and its target genes can be potential biomarkers for diagnosis of PTL.
Highlights
The placenta, a maternal–fetal interface organ, is important for fetal development and protection and maintenance of pregnancy, because various pregnancy-related hormones, growth factors, nutrients and waste products are transported through the placenta [1]
Preterm labor occurs due to several obstacles such as inflammation as well as blood supply into the placenta, as trophoblast cells are not penetrated into maternal portion [19]
We found that miR-373-3p simultaneously target two genes, CD44 and RDX, through the combination between miRNA array and gene array in placental tissues and trophoblast cells
Summary
The placenta, a maternal–fetal interface organ, is important for fetal development and protection and maintenance of pregnancy, because various pregnancy-related hormones, growth factors, nutrients and waste products are transported through the placenta [1]. It is necessary to study the potential function of miR-373-3p and the miRNA-target genes in the migration and invasion of trophoblast cells, which will help to understand mechanisms of implantation and placental development. MiRNA-373-3p Targets Two Genes, CD44 and RDX at the Same Manner in Placenta As Well As Trophoblast CellTso validate the expression of miR-373-3p, we conducted qRT-PCR from RNA in the placental tissues. In localization oaf nCeDga4t4ivaendcoRnDtroXl,ininptelraecsetnintagllyti,stshueese,xwpreescsoinodnuocfteRdDiXmamsuwneolflluaosrCesDce4n4cwe.as gradually As shown irneduFcigeudrien t2hGe,orCdeDr4o4f i(ngflraemenm) aatonrdy tRerDmXan(drePdT) Lwpelarecenetxaps,ressimseidlaritno tthheeWestern blot non-inflammraetsourlytst.eTrmhispmlaeceantsatlhtaistseuxep.rIenssaionnegoaf tCivDe4c4oanntrdoRl, DinXteirsemstionrgeldy,ecthreeaesxedpriens-PTL placenta sion of RDX tahsawn etellrmaspClaDce4n4tawwasitghraodr uwailtlhyoruetdiuncfleadmimn athtieono.rder of inflammatory term and PTL placentas, similar to the Western blot results This means that expression of CD44 and RDX is more decreased in PTL placenta than term placenta with or without inflammation. EAxspwresesicoonnofifrmmieRd-3t7h3e-3epx,pCreDs4si4oannodfRmDiXR-i3n73N-o3rpmaanl danCdDP4T4LaPnadtieRnDtsX’ Binlotoedrm and PTL placenAtsawtiesscuoensfi, rwmeecdotnhdeuecxtepdreqssRioTn-PoCfRmaiRna-3ly7s3i-s3ptoavnadliCdDat4e4thaneidr RexDpXreisnsitoenrminacnldinPicTaLl npolarmceanltaantidssPuTeLs,pwaetiecnotnsd. umcitRed-37q3R-T3-pPiCsRmaonraeliynscisretaosevdaliindaPtTeLthpeairtieexnptsr’ebssloioonditnhaclnintihcaatl onfonrmoraml aanl dpaPtTieLntpsa,taienndtsi.tsmeiRxp-3r7e3ss-3iopnisismmoruecihncmreoarseedinicnrePaTsLedpabtyieinntfsl’abmlomoadtitohnan(Fthigaut roef 6nAo)r.mBayl tpaartgieetnintsg, amnidRi-t3s7e3x-3ppre, sCsDio4n4iasnmduRchDXmodreecrienacsreeamseodrebiyninPflTaLmpmataietinotns’(bFliogoudreth6aAn). iBnythtaartgoeftinnogrmmiaRl-p37a3ti-e3npt,sC, aDn4d4 tahnedirReDxpXrdesesciroenasies mmoorreeidnePcTreLapseadtiebnytsi’nbfllaomodmtahtainonin(Fthiga-t uorfen6oBrm,Ca).l pTahteiseentrse,saunldtstchoeiinrceixdperwesistihonreissumltos rwe idthecprelaacseedntbaytiisnsfluaems (mFiagtiuorne (2FDig,Eu)reex6cBe,Cpt). fTohreCseDr4e4s,ualtnsdcoiitnrceipdreewseinthtsrethsueltvsawliditahtepdlacreensutalttsiswsuitehs (cFliingiucrael 2pDa,tEie)netxsc’ebpltofoodr CasDw44e,llanads pitlarceepnrteasetnistssutehse. vTahliedraeftoedrer,eisturletspwreistehnctslinthicealppoatetinetniatsl’sbolof omdiRas-3w73e-ll3pasapslaacesnpteacitfiiscsubeis-. oTmhearrekfeorrew, hiticrhepcroeusledntdsettheectpPoTteLn. tials of miR-373-3p as a specific biomarker which could detect PTL
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