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Pregnancy-associated microRNAs in plasma as potential molecular markers of ectopic pregnancy

      Objective

      To investigate cell-free pregnancy-associated microRNAs as molecular markers for the diagnosis of ectopic pregnancy.

      Design

      Laboratory study using human plasma samples.

      Setting

      Research unit in a university hospital.

      Patient(s)

      Plasma samples from 18 women with ectopic pregnancies (EP group), 12 women with spontaneous abortion (SA group), and 26 normal women with singleton pregnancies (NP group).

      Intervention(s)

      Total RNAs containing small RNA molecules extracted from 1.2 mL of plasma.

      Main Outcome Measure(s)

      Plasma concentrations of cell-free microRNAs measured by quantitative real-time reverse-transcriptase polymerase chain reaction.

      Result(s)

      Plasma concentrations of cell-free pregnancy-associated microRNAs (miR-323-3p, miR-515-3p, miR-517a, miR-517c, and miR-518b) and serum concentration of human chorionic gonadotropin (hCG) were confirmed to have statistically significantly different plasma or serum concentrations in women with EP, SA, or NP. There was no statistically significant difference in the plasma concentrations of cell-free miR-21 between the three groups. By correlation coefficient analysis, no relationship was detected between serum hCG levels and plasma cell-free miR-517c, miR-515-3p, miR-517a, miR-518b, miR-323-3p, or miR-21 levels. Plasma concentrations of cell-free miR-517a could distinguish EP/SA from NP, yielding an area under the curve of 0.9654 (95% confidence interval, 0.9172–1.0). Plasma concentrations of cell-free miR-323-3p could distinguish EP from SA, yielding an area under the curve of 0.7454 (95% confidence interval, 0.5558–0.9349).

      Conclusion(s)

      Cell-free pregnancy-associated microRNAs have potential as molecular markers of ectopic pregnancy.

      Key Words

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      References

        • Barnhart K.T.
        Clinical practice. Ectopic pregnancy.
        N Engl J Med. 2009; 361: 379-387
        • Farquhar C.M.
        Ectopic pregnancy.
        Lancet. 2005; 366: 583-591
        • American College of Obstetricians Gynecologists
        ACOG Practice Bulletin No. 94: medical management of ectopic pregnancy.
        Obstet Gynecol. 2008; 111: 1479-1485
        • Zhao Z.
        • Zhao Q.
        • Warrick J.
        • Lockwood C.M.
        • Woodworth A.
        • Moley K.H.
        • et al.
        Circulating microRNA miR-323-3p as a biomarker of ectopic pregnancy.
        Clin Chem. 2012; 58: 896-905
        • Chim S.S.
        • Shing T.K.
        • Hung E.C.
        • Leung T.Y.
        • Lau T.K.
        • Chiu R.W.
        • et al.
        Detection and characterization of placental microRNAs in maternal plasma.
        Clin Chem. 2008; 54: 482-490
        • Chiu R.W.
        • Lo Y.M.
        Pregnancy-associated microRNAs in maternal plasma: a channel for fetal-maternal communication?.
        Clin Chem. 2010; 56: 1656-1657
        • Miura K.
        • Miura S.
        • Yamasaki K.
        • Higashijima A.
        • Kinoshita A.
        • Yoshiura K.
        • et al.
        Identification of pregnancy-associated microRNAs in maternal plasma.
        Clin Chem. 2010; 56: 1767-1771
        • Ng E.K.
        • Tsui N.B.
        • Lau T.K.
        • Leung T.N.
        • Chiu R.W.
        • Panesar N.S.
        • et al.
        mRNA of placental origin is readily detectable in maternal plasma.
        Proc Natl Acad Sci U S A. 2003; 100: 4748-4753
        • Heung M.M.
        • Jin S.
        • Tsui N.B.
        • Ding C.
        • Leung T.Y.
        • Lau T.K.
        • et al.
        Placenta-derived fetal specific mRNA is more readily detectable in maternal plasma than in whole blood.
        PLoS One. 2009; 4: e5858
        • Chiu R.W.
        • Poon L.L.
        • Lau T.K.
        • Leung T.N.
        • Wong E.M.
        • Lo Y.M.
        Effects of blood-processing protocols on fetal and total DNA quantification in maternal plasma.
        Clin Chem. 2001; 47: 1607-1613
        • Donker R.B.
        • Mouillet J.F.
        • Chu T.
        • Hubel C.A.
        • Stolz D.B.
        • Morelli A.E.
        • et al.
        The expression profile of C19MC microRNAs in primary human trophoblast cells and exosomes.
        Mol Hum Reprod. 2012; 18: 417-424
        • Luo S.S.
        • Ishibashi O.
        • Ishikawa G.
        • Ishikawa T.
        • Katayama A.
        • Mishima T.
        • et al.
        Human villous trophoblasts express and secrete placenta-specific microRNAs into maternal circulation via exosomes.
        Biol Reprod. 2009; 81: 717-729
        • Higashijima A.
        • Miura K.
        • Mishima H.
        • Kinoshita A.
        • Jo O.
        • Abe S.
        • et al.
        Characterization of placenta-specific microRNAs in fetal growth restriction pregnancy.
        Prenat Diagn. 2013; 33: 214-222
        • Hasegawa Y.
        • Miura K.
        • Furuya K.
        • Yoshiura K.I.
        • Masuzaki H.
        Identification of complete hydatidiform mole pregnancy-associated microRNAs in plasma.
        Clin Chem. 2013; 59: 1410-1412
        • Ng E.K.
        • Tsui N.B.
        • Lam N.Y.
        • Chiu R.W.
        • Yu S.C.
        • Wong S.C.
        • et al.
        Presence of filterable and nonfilterable mRNA in the plasma of cancer patients and healthy individuals.
        Clin Chem. 2002; 48: 1212-1217
        • Robin X.
        • Turck N.
        • Hainard A.
        • Tiberti N.
        • Lisacek F.
        • Sanchez J.-C.
        • et al.
        pROC: an open-source package for R and S+ to analyze and compare ROC curves.
        BMC Bioinformatics. 2011; 12: 77
        • Noguer-Dance M.
        • Abu-Amero S.
        • Al-Khtib M.
        • Lefèvre A.
        • Coullin P.
        • Moore G.E.
        • et al.
        The primate-specific microRNA gene cluster (C19MC) is imprinted in the placenta.
        Hum Mol Genet. 2010; 19: 3566-3582
        • Lee R.C.
        • Feinbaum R.L.
        • Ambros V.
        The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14.
        Cell. 1993; 75: 843-854
        • Mendell J.T.
        MicroRNAs: critical regulators of development, cellular physiology and malignancy.
        Cell Cycle. 2005; 4: 1179-1184
        • Plasterk R.H.
        Micro RNAs in animal development.
        Cell. 2006; 124: 877-881
        • Lozoya T.
        • Domínguez F.
        • Romero-Ruiz A.
        • Steffani L.
        • Martínez S.
        • Monterde M.
        • et al.
        The Lin28/Let-7 system in early human embryonic tissue and ectopic pregnancy.
        PLoS One. 2014; 9: e87698
        • Morales-Prieto D.M.
        • Ospina-Prieto S.
        • Chaiwangyen W.
        • Schoenleben M.
        • Markert U.R.
        Pregnancy-associated miRNA-clusters.
        J Reprod Immunol. 2013; 97: 51-61
        • Galliano D.
        • Pellicer A.
        MicroRNA and implantation.
        Fertil Steril. 2014; 101: 1531-1544