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Short communication| Volume 33, P161-167, March 2018

Semen-specific miRNAs: Suitable for the distinction of infertile semen in the body fluid identification?

  • Author Footnotes
    1 These authors contributed equally to this work.
    Huan Tian
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Department of Forensic Genetics, West China School of Basic Medical Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China
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  • Author Footnotes
    1 These authors contributed equally to this work.
    Meili Lv
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Department of Forensic Genetics, West China School of Basic Medical Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China
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  • Zhilong Li
    Affiliations
    Department of Forensic Genetics, West China School of Basic Medical Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China
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  • Duo Peng
    Affiliations
    Department of Forensic Genetics, West China School of Basic Medical Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China
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  • Yu Tan
    Affiliations
    Department of Forensic Genetics, West China School of Basic Medical Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China
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  • Hui Wang
    Affiliations
    Institute of Forensic Science, Chengdu Public Security Bureau, Chengdu 610081, Sichuan, China
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  • Qingqing Li
    Affiliations
    Institute of Forensic Science, Chengdu Public Security Bureau, Chengdu 610081, Sichuan, China
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  • Fuping Li
    Correspondence
    Corresponding authors.
    Affiliations
    Human Sperm Bank, Key Laboratory of Birth Defects and Related Diseases of Women and Children of Ministry of Education, West China Second University Hospital of Sichuan University, Chengdu, 610041, Sichuan, China
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  • Weibo Liang
    Correspondence
    Corresponding authors.
    Affiliations
    Department of Forensic Genetics, West China School of Basic Medical Science and Forensic Medicine, Sichuan University, Chengdu, 610041, Sichuan, China
    Search for articles by this author
  • Author Footnotes
    1 These authors contributed equally to this work.
Published:December 19, 2017DOI:https://doi.org/10.1016/j.fsigen.2017.12.010

      Highlights

      • The specificity of 6 miRNAs were validated and confirmed in normal semen samples.
      • The expression of 6 semen-specific miRNAs were detected in infertile semen samples.
      • The effect of infertile semen samples on BFI were further evaluated by two-dimensional scatter plots and discriminant function.
      • The results of our study provide guides and references for the application of miRNAs in forensic body fluid identification.

      Abstract

      Non-protein coding RNA, miRNAs (microRNAs), are a class of promising molecular biomarkers for forensic body fluid identification (BFI) as their small size and tissue-specific expression manners. A number of studies have shown that semen can be distinguished from forensic-related body fluids (such as menstrual blood, venous blood, vaginal fluid, saliva, etc.) using semen-specific miRNAs through microassay screening and RT-qRCR. Infertility is becoming a global health problem, affecting 10%-15% of couples worldwide, and half of the cases are the result of male factors (Lian et al., 2009 [
      • Lian J.
      • Zhang X.
      • Tian H.
      • Liang N.
      • Wang Y.
      • Liang C.
      • Li X.
      • Sun F.
      Altered microRNA expression in patients with non-obstructive azoospermia.
      ]). Forensic researchers have to consider the impact of semen infertility on semen identification with a high incidence of infertility. In the present study, normal semen (NS) and four other types of infertile semen samples, including asthenospermia (AS), oligospermia (OS), azoospermia (AZ), oligospermia and asthenospermia (OSAS) semen, were collected. The expression levels of a set of semen-specific miRNA markers (miR-10a, miR-10b, miR-135a, miR-135b, miR-888 and miR-891a) were evaluated using a real-time quantitative PCR technique with a specific fluorescence-labelled TaqMan probe. The results showed the significantly high expression of these miRNAs in normal semen, and the molecules have semen specificity. Nevertheless, a distinct down-regulation in the expression of infertile samples compared with normal semen samples was observed. Moreover, differences in the results of selected optimal biomarkers between the discriminant function and two-dimensional scatter plots were also detected. The goal of the present study was to identify a small set of semen-specific miRNAs that efficiently and accurately distinguish semen (fertile and infertile) from other forensic-related body fluids. The results of the present study suggest that attention should be paid to infertile semen samples when using miRNAs to identify semen samples, for which would have a far-reaching impact on forensic identification.

      Keywords

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      References

        • Lian J.
        • Zhang X.
        • Tian H.
        • Liang N.
        • Wang Y.
        • Liang C.
        • Li X.
        • Sun F.
        Altered microRNA expression in patients with non-obstructive azoospermia.
        Reprod. Biol. Endocrinol. 2009; 7: 13
        • Hanson E.K.
        • Lubenow H.
        • Ballantyne J.
        Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs.
        Anal. Biochem. 2009; 387: 303-314
        • Wang Z.
        • Zhang J.
        • Luo H.
        • Ye Y.
        • Yan J.
        • Hou Y.
        Screening and confirmation of microRNA markers for forensic body fluid identification.
        Forensic Sci. Int. Genet. 2013; 7: 116-123
        • Silva S.S.
        • Lopes C.
        • Teixeira A.L.
        • Carneiro de Sousa M.J.
        • Medeiros R.
        Forensic miRNA: potential biomarker for body fluids?.
        Forensic Sci. Int. Genet. 2015; 14: 1-10
        • Sauer E.
        • Reinke A.K.
        • Courts C.
        Differentiation of five body fluids from forensic samples by expression analysis of four microRNAs using quantitative PCR.
        Forensic Sci. Int. Genet. 2016; 22: 89-99
        • Tobe S.S.
        • Watson N.
        • Daeid N.N.
        Evaluation of six presumptive tests for blood, their specificity, sensitivity, and effect on high molecular-weight DNA.
        J. Forensic Sci. 2007; 52: 102-109
        • Myers J.R.
        • Adkins W.K.
        Comparison of modern techniques for saliva screening.
        J. Forensic Sci. 2008; 53: 862-867
        • Khaldi N.
        • Miras A.
        • Botti K.
        • Benali L.
        • Gromb S.
        Evaluation of three rapid detection methods for the forensic identification of seminal fluid in rape cases.
        J. Forensic Sci. 2004; 49: 749-753
        • Park J.L.
        • Park S.M.
        • Kwon O.H.
        • Lee H.C.
        • Kim J.Y.
        • Seok H.H.
        • Lee W.S.
        • Lee S.H.
        • Kim Y.S.
        • Woo K.M.
        • Kim S.Y.
        Microarray screening and qRT-PCR evaluation of microRNA markers for forensic body fluid identification.
        Electrophoresis. 2014; 35: 3062-3068
        • Bartel D.P.
        MicroRNAs: genomics, biogenesis, mechanism, and function.
        Cell. 2004; 116: 281-297
        • Hayashi K.
        • Chuva de Sousa Lopes S.M.
        • Kaneda M.
        • Tang F.
        • Hajkova P.
        • Lao K.
        • O'Carroll D.
        • Das P.P.
        • Tarakhovsky A.
        • Miska E.A.
        • Surani M.A.
        MicroRNA biogenesis is required for mouse primordial germ cell development and spermatogenesis.
        PLoS One. 2008; 3: e1738
        • Ambros V.
        The functions of animal microRNAs.
        Nature. 2004; 431: 350-355
        • Bartel D.P.
        MicroRNAs: target recognition and regulatory functions.
        Cell. 2009; 136: 215-233
        • Wu W.
        • Qin Y.
        • Li Z.
        • Dong J.
        • Dai J.
        • Lu C.
        • Guo X.
        • Zhao Y.
        • Zhu Y.
        • Zhang W.
        • Hang B.
        • Sha J.
        • Shen H.
        • Xia Y.
        • Hu Z.
        • Wang X.
        Genome-wide microRNA expression profiling in idiopathic non-obstructive azoospermia: significant up-regulation of miR-141 miR-429 and miR-7-1-3p.
        Hum. Reprod. 2013; 28: 1827-1836
        • Zubakov D.
        • Boersma A.M.
        • Choi Y.
        • van Kuijk P.
        • Wiemer E.C.
        • Kayser M.
        MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation.
        Int. J. Legal Med. 2010; 124: 217-226
        • Ruan J.
        • He X.J.
        • Du W.D.
        • Chen G.
        • Zhou Y.
        • Xu S.
        • Zuo X.B.
        • Fang L.B.
        • Cao Y.X.
        • Zhang X.J.
        Genetic variants in TEX15 gene conferred susceptibility to spermatogenic failure in the Chinese Han population.
        Reprod. Sci. 2012; 19: 1190-1196
        • Okada H.
        • Tajima A.
        • Shichiri K.
        • Tanaka A.
        • Tanaka K.
        • Inoue I.
        Genome-wide expression of azoospermia testes demonstrates a specific profile and implicates ART3 in genetic susceptibility.
        PLoS Genet. 2008; 4: e26
        • Hirsh A.
        Male subfertility.
        BMJ. 2003; 327: 669-672
        • Vogt P.H.
        Molecular genetics of human male infertility: from genes to new therapeutic perspectives.
        Curr. Pharm. Des. 2004; 10: 471-500
        • Wu W.
        • Hu Z.
        • Qin Y.
        • Dong J.
        • Dai J.
        • Lu C.
        • Zhang W.
        • Shen H.
        • Xia Y.
        • Wang X.
        Seminal plasma microRNAs: potential biomarkers for spermatogenesis status.
        Mol. Hum. Reprod. 2012; 18: 489-497
        • Abu-Halima M.
        • Hammadeh M.
        • Schmitt J.
        • Leidinger P.
        • Keller A.
        • Meese E.
        • Backes C.
        Altered microRNA expression profiles of human spermatozoa in patients with different spermatogenic impairments.
        Fertil. Steril. 2013; 1255: e1216
        • Cooper T.G.
        • Noonan E.
        • von Eckardstein S.
        • Auger J.
        • Baker H.W.
        • Behre H.M.
        • Haugen T.B.
        • Kruger T.
        • Wang C.
        • Mbizvo M.T.
        • Vogelsong K.M.
        World Health Organization reference values for human semen characteristics.
        Hum. Reprod. Update. 2010; 16: 231-245
        • Sauer E.
        • Madea B.
        • Courts C.
        An evidence based strategy for normalization of quantitative PCR data from miRNA expression analysis in forensically relevant body fluids.
        Forensic Sci. Int. Genet. 2014; 11: 174-181
        • Sauer E.
        • Babion I.
        • Madea B.
        • Courts C.
        An evidence based strategy for normalization of quantitative PCR data from miRNA expression analysis in forensic organ tissue identification.
        Forensic Sci. Int. Genet. 2014; 13: 217-223
        • Tong D.
        • Jin Y.
        • Xue T.
        • Ma X.
        • Zhang J.
        • Ou X.
        • Cheng J.
        • Sun H.
        Investigation of the Application of miR10b and miR135b in the Identification of Semen Stains.
        PLoS One. 2015; 10: e0137067
        • Seashols-Williams S.
        • Lewis C.
        • Calloway C.
        • Peace N.
        • Harrison A.
        • Hayes-Nash C.
        • Fleming S.
        • Wu Q.
        • Zehner Z.E.
        High-throughput miRNA sequencing and identification of biomarkers for forensically relevant biological fluids.
        Electrophoresis. 2016;
        • Hu L.
        • Wu C.
        • Guo C.
        • Li H.
        • Xiong C.
        Identification of microRNAs predominately derived from testis and epididymis in human seminal plasma.
        Clin. Biochem. 2014; 47: 967-972
        • Li Z.
        • Bai P.
        • Peng D.
        • Wang H.
        • Guo Y.
        • Jiang Y.
        • He W.
        • Tian H.
        • Yang Y.
        • Huang Y.
        • Long B.
        • Liang W.
        • Zhang L.
        Screening and confirmation of microRNA markers for distinguishing between menstrual and peripheral blood.
        Forensic Sci. Int. Genet. 2017; 30: 24-33
        • Zubakov D.
        • Hanekamp E.
        • Kokshoorn M.
        • van W.
        • Jcken I.
        • Kayser M.
        Stable RNA markers for identification of blood and saliva stains revealed from whole genome expression analysis of time-wise degraded samples.
        Int. J. Legal Med. 2008; 122: 135-142
        • Weber J.A.
        • Baxter D.H.
        • Zhang S.
        • Huang D.Y.
        • Huang K.H.
        • Lee M.J.
        • Galas D.J.
        • Wang K.
        The microRNA spectrum in 12 body fluids.
        Clin. Chem. 2010; 56: 1733-1741
        • Zubakov D.
        • Kokshoorn M.
        • Kloosterman A.
        • Kayser M.
        New markers for old stains: stable mRNA markers for blood and saliva identification from up to 16-year-old stains.
        Int. J. Legal Med. 2009; 123: 71-74
        • Haas C.
        • Hanson E.
        • Anjos M.J.
        • Banemann R.
        • Berti A.
        • Borges E.
        • Carracedo A.
        • Carvalho M.
        • Courts C.
        • De Cock G.
        • Dotsch M.
        • Flynn S.
        • Gomes I.
        • Hollard C.
        • Hjort B.
        • Hoff-Olsen P.
        • Hribikova K.
        • Lindenbergh A.
        • Ludes B.
        • Maronas O.
        • McCallum N.
        • Moore D.
        • Morling N.
        • Niederstatter H.
        • Noel F.
        • Parson W.
        • Popielarz C.
        • Rapone C.
        • Roeder A.D.
        • Ruiz Y.
        • Sauer E.
        • Schneider P.M.
        • Sijen T.
        • Court D.S.
        • Sviezena B.
        • Turanska M.
        • Vidaki A.
        • Zatkalikova L.
        • Ballantyne J.
        RNA/DNA co-analysis from human saliva and semen stains--results of a third collaborative EDNAP exercise.
        Forensic Sci. Int. Genet. 2013; 7: 230-239
        • Hochmeister M.N.
        • Budowle B.
        • Rudin O.
        • Gehrig C.
        • Borer U.
        • Thali M.
        • Dirnhofer R.
        Evaluation of prostate-specific antigen (PSA) membrane test assays for the forensic identification of seminal fluid.
        J. Forensic Sci. 1999; 44: 1057-1060
        • van Rooij E.
        (99)The art of MicroRNA research.
        Circ. Res. 2011; 108: 219-234
        • Lehmann S.M.
        • Kruger C.
        • Park B.
        • Derkow K.
        • Rosenberger K.
        • Baumgart J.
        • Trimbuch T.
        • Eom G.
        • Hinz M.
        • Kaul D.
        • Habbel P.
        • Kalin R.
        • Franzoni E.
        • Rybak A.
        • Nguyen D.
        • Veh R.
        • Ninnemann O.
        • Peters O.
        • Nitsch R.
        • Heppner F.L.
        • Golenbock D.
        • Schott E.
        • Ploegh H.L.
        • Wulczyn F.G.
        • Lehnardt S.
        An unconventional role for miRNA: let-7 activates Toll-like receptor 7 and causes neurodegeneration.
        Nat. Neurosci. 2012; 15: 827-835
        • Hébert S.S.
        • De B.
        Strooper, miRNAs in neurodegeneration.
        Science. 2007; 317: 1179-1180
        • Abe M.
        • Bonini N.M.
        (96)MicroRNAs and neurodegeneration: role and impact.
        Trends Cell Biol. 2016; 23: 30-36
        • Nelson P.T.
        • Wang W.X.
        • Rajeev B.W.
        MicroRNAs (miRNAs) in neurodegenerative diseases.
        Brain Pathol. 2008; 18: 130-138
        • Welberg L.
        Neurodegenerative disorders: reconnect with microRNA.
        Nat. Rev. Neurosci. 2010; 11: 74
        • Courts C.
        • Madea B.
        Specific micro-RNA signatures for the detection of saliva and blood in forensic body-fluid identification.
        J. Forensic Sci. 2011; 56: 1464-1470
        • Fayyad-Kazan H.
        • Bitar N.
        • Najar M.
        • Lewalle P.
        • Fayyad-Kazan M.
        • Badran R.
        • Hamade E.
        • Daher A.
        • Hussein N.
        • ElDirani R.
        • Berri F.
        • Vanhamme L.
        • Burny A.
        • Martiat P.
        • Rouas R.
        • Badran B.
        Circulating miR-150 and miR-342 in plasma are novel potential biomarkers for acute myeloid leukemia.
        J. Transl. Med. 2013; 11: 31
        • Hanke M.
        • Hoefig K.
        • Merz H.
        • Feller A.C.
        • Kausch I.
        • Jocham D.
        • Warnecke J.M.
        • Sczakiel G.
        A robust methodology to study urine microRNA as tumor marker: microRNA-126 and microRNA-182 are related to urinary bladder cancer.
        Urol. Oncol. 2010; 28: 655-661