title: Deep-sequencing identification of differentially expressed miRNAs in decidua and villus of recurrent miscarriage patients.
abstract: OBJECTIVE
  MicroRNAs (miRNAs) are small non-coding RNA molecules that play critical roles in post-transcriptional gene expression regulation. The aim of this study was to identify differentially expressed miRNAs in decidua and villus of recurrent miscarriage (RM) patients.
  
  
  METHODS
  Participants were recruited at the outpatient Department of Gynecology and Obstetrics, The Second Hospital of Tianjin Medical University, China. Decidua and villus tissues were collected by curettage from recruited RM patients and normal pregnant women with their informed consent. MiRNAs expression profiles in decidua or villus were respectively determined by the deep-sequencing analysis. The predicated target genes of these differentially expressed miRNAs were analyzed by miRWalk. The differential expressions of four miRNAs in decidua and four miRNAs in villus between the six pairs of RM patients and normal pregnant women were confirmed by qRT-PCR analysis. The expression patterns of two predicated target genes, Bcl-2 and Pten, in the same six pairs of decidual or villus tissues were detected by Western blotting analysis, respectively.
  
  
  RESULTS
  Totally 18 RM patients and 15 normal pregnant women were recruited. Thirty-two miRNAs in decidua and four miRNAs in villus of RM patients were screened out to be significantly up-regulated compared to that of normal pregnant women, and five miRNAs in villus of RM patients were screened out to be remarkably down-regulated compared to that of normal pregnant women (P value < 0.05 and Fold change >2). These differentially expressed miRNAs were predicted to target a large number of genes that involved in cell apoptosis, p53 signaling pathway, cell cycle and other cellular bio-functions. Differential expressions of hsa-miR-516a-5p, -517a-3p, -519a-3p and -519d in decidua, as well as hsa-miR-1, -372, -100-5p and -146a-5p in villus, were validated by qRT-PCR analysis. In the decidual of RM patients, expression of hsa-miR-516a-5p, -517a-3p, -519a-3p and -519d were significantly up-regulated compared to normal pregnancy. In the villi of RM patients, expression of hsa-miR-100 and -146a-5p were significantly higher, while hsa-miR-1 and -372 were significantly lower compared to normal pregnancy. Furthermore, the expression of Bcl-2 and Pten, a predicated target gene of hsa-miR-1 or hsa-miR-372 respectively, was significantly up-regulated in the villi of RM patients.
  
  
  CONCLUSIONS
  These data suggested that the pathogenic process of RM might be associated with the alteration of miRNAs expression profiles in decidua and villus. Especially, the aberrant placental expression of hsa-miR-1 and -372 might be involved in the progression of RM, but need to be further investigated by larger studies in the future.
authors.affiliation: The Second Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Jian-Mei
authors.initials: JM
authors.lastname: Wang
authors.affiliation: The Second Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gu
authors.affiliation: Institute of Biochemistry and Cell Biology, SIBS, CAS, Shanghai, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: NPFPC Key Laboratory of Contraceptive Drugs and Devices, Shanghai Institute of Planned Parenthood Research, 2140 Xie Tu Road, 200032, Shanghai, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Yang
authors.affiliation: NPFPC Key Laboratory of Contraceptive Drugs and Devices, Shanghai Institute of Planned Parenthood Research, 2140 Xie Tu Road, 200032, Shanghai, China. xuanzhang@sippr.org.cn.
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhang
authors.affiliation: The Second Hospital of Tianjin Medical University, Tianjin, China.
authors.forename: Lirong
authors.initials: L
authors.lastname: Yin
authors.affiliation: NPFPC Key Laboratory of Contraceptive Drugs and Devices, Shanghai Institute of Planned Parenthood Research, 2140 Xie Tu Road, 200032, Shanghai, China. wangjian@sippr.org.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
mesh_terms: D000026:Abortion, Habitual / Q000235:genetics*; D000328:Adult; D002681:China; D002824:Chorionic Villi / Q000737:chemistry / Q000378:metabolism*; D003656:Decidua / Q000737:chemistry / Q000378:metabolism*; D015536:Down-Regulation; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D005819:Genetic Markers; D006801:Humans; D035683:MicroRNAs / Q000032:analysis*; D010920:Placenta / Q000737:chemistry* / Q000473:pathology / Q000503:physiopathology; D011247:Pregnancy; D015854:Up-Regulation
chemical_list: D005819:Genetic Markers; C544547:MIRN1 microRNA, human; C544266:MIRN100 microRNA, human; C561021:MIRN372 microRNA, human; D035683:MicroRNAs
keywords: Decidua; Deep-sequencing; Recurrent miscarriage; Villus; miRNAs