Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Journal of Assisted Reproduction and Genetics
Published in: Journal of Assisted Reproduction and Genetics 1/2016

01-01-2016 | Reproductive Physiology and Disease

Aberrant expression of long noncoding RNAs in cumulus cells isolated from PCOS patients

Authors: Xin Huang, Cuifang Hao, Hongchu Bao, Meimei Wang, Huangguan Dai

Published in: Journal of Assisted Reproduction and Genetics | Issue 1/2016

Login to get access

Abstract

Purpose

To describe the long noncoding RNA (lncRNA) profiles in cumulus cells isolated from polycystic ovary syndrome (PCOS) patients by employing a microarray and in-depth bioinformatics analysis. This information will help us understand the occurrence and development of PCOS.

Methods

In this study, we used a microarray to describe lncRNA profiles in cumulus cells isolated from ten patients (five PCOS and five normal women). Several differentially expressed lncRNAs were chosen to validate the microarray results by quantitative RT-PCR (qRT-PCR). Then, the differentially expressed lncRNAs were classified into three subgroups (HOX loci lncRNA, enhancer-like lncRNA, and lincRNA) to deduce their potential features. Furthermore, a lncRNA/mRNA co-expression network was constructed by using the Cytoscape software (V2.8.3, http://​www.​cytoscape.​org/​).

Results

We observed that 623 lncRNAs and 260 messenger RNAs (mRNAs) were significantly up- or down-regulated (≥2-fold change), and these differences could be used to discriminate cumulus cells of PCOS from those of normal patients. Five differentially expressed lncRNAs (XLOC_011402, ENST00000454271, ENST00000433673, ENST00000450294, and ENST00000432431) were selected to validate the microarray results using quantitative RT-PCR (qRT-PCR). The qRT-PCR results were consistent with the microarray data. Further analysis indicated that many differentially expressed lncRNAs were transcribed from chromosome 2 and may act as enhancers to regulate their neighboring protein-coding genes. Forty-three lncRNAs and 29 mRNAs were used to construct the coding-non-coding gene co-expression network. Most pairs positively correlated, and one mRNA correlated with one or more lncRNAs.

Conclusions

Our study is the first to determine genome-wide lncRNA expression patterns in cumulus cells isolated from PCOS patients by microarray. The results show that clusters of lncRNAs were aberrantly expressed in cumulus cells of PCOS patients compared with those of normal women, which revealed that lncRNAs differentially expressed in PCOS and normal women may contribute to the occurrence of PCOS and affect oocyte development.
Appendix
Available only for authorised users
Literature
1.
Shi Y, Zhao H, Cao Y, Yang D, Li Z, Zhang B, et al. Genome-wide association study identifies eight new risk loci for polycystic ovary syndrome. Nat Genet. 2012;44(9):1020–5.PubMedCrossRef
2.
Diao FY, Xu M, Hu Y, Li J, Xu Z, Lin M, et al. The molecular characteristics of polycystic ovary syndrome (PCOS) ovary defined by human ovary cDNA microarray. J Mol Endocrinol. 2004;33(1):59–72.PubMedCrossRef
3.
Group REA-SPCW. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome. Fertil Steril. 2004;81(1):19–25.CrossRef
4.
Kandaraki E, Christakou C, Diamanti-Kandarakis E. Metabolic syndrome and polycystic ovary syndrome… vice versa. Arq Bras Endocrinol Metabol. 2009;53(2):227–37.PubMedCrossRef
5.
Carmina E. Cardiovascular risk and events in polycystic ovary syndrome. Climacteric. 2009;12 Suppl 1:22–5.PubMedCrossRef
6.
Wild S, Pierpoint T, Jacobs H, McKeigue P. Long-term consequences of polycystic ovary syndrome: results of a 31 year follow-up study. Hum Fertil (Camb). 2000;3(2):101–5.CrossRef
7.
Diamanti-Kandarakis E, Piperi C. Genetics of polycystic ovary syndrome: searching for the way out of the labyrinth. Hum Reprod Update. 2005;11(6):631–43.PubMedCrossRef
8.
Vink JM, Sadrzadeh S, Lambalk CB, Boomsma DI. Heritability of polycystic ovary syndrome in a Dutch twin-family study. J Clin Endocrinol Metab. 2006;91(6):2100–4.PubMedCrossRef
9.
Wood JR, Ho CK, Nelson-Degrave VL, McAllister JM, Strauss 3rd JF. The molecular signature of polycystic ovary syndrome (PCOS) theca cells defined by gene expression profiling. J Reprod Immunol. 2004;63(1):51–60.PubMedCrossRef
10.
Jansen E, Laven JS, Dommerholt HB, Polman J, van Rijt C, van den Hurk C, et al. Abnormal gene expression profiles in human ovaries from polycystic ovary syndrome patients. Mol Endocrinol. 2004;18(12):3050–63.PubMedCrossRef
11.
Oksjoki S, Soderstrom M, Inki P, Vuorio E, Anttila L. Molecular profiling of polycystic ovaries for markers of cell invasion and matrix turnover. Fertil Steril. 2005;83(4):937–44.PubMedCrossRef
12.
Wood JR, Dumesic DA, Abbott DH, Strauss 3rd JF. Molecular abnormalities in oocytes from women with polycystic ovary syndrome revealed by microarray analysis. J Clin Endocrinol Metab. 2007;92(2):705–13.PubMedCrossRef
13.
Kenigsberg S, Bentov Y, Chalifa-Caspi V, Potashnik G, Ofir R, Birk OS. Gene expression microarray profiles of cumulus cells in lean and overweight-obese polycystic ovary syndrome patients. Mol Hum Reprod. 2009;15(2):89–103.PubMedCrossRef
14.
Mohamed-Hussein ZA, Harun S. Construction of a polycystic ovarian syndrome (PCOS) pathway based on the interactions of PCOS-related proteins retrieved from bibliomic data. Theor Biol Med Model. 2009;6:18.PubMedPubMedCentralCrossRef
15.
Ponting CP, Oliver PL, Reik W. Evolution and functions of long noncoding RNAs. Cell. 2009;136(4):629–41.PubMedCrossRef
16.
17.
Roth LW, McCallie B, Alvero R, Schoolcraft WB, Minjarez D, Katz-Jaffe MG. Altered microRNA and gene expression in the follicular fluid of women with polycystic ovary syndrome. J Assist Reprod Genet. 2014;31(3):355–62.PubMedPubMedCentralCrossRef
18.
Sang Q, Yao Z, Wang H, Feng R, Zhao X, Xing Q, et al. Identification of microRNAs in human follicular fluid: characterization of microRNAs that govern steroidogenesis in vitro and are associated with polycystic ovary syndrome in vivo. J Clin Endocrinol Metab. 2013;98(7):3068–79.PubMedCrossRef
19.
Moran I, Akerman I, van de Bunt M, Xie R, Benazra M, Nammo T, et al. Human beta cell transcriptome analysis uncovers lncRNAs that are tissue-specific, dynamically regulated, and abnormally expressed in type 2 diabetes. Cell Metab. 2012;16(4):435–48.PubMedPubMedCentralCrossRef
20.
Klattenhoff CA, Scheuermann JC, Surface LE, Bradley RK, Fields PA, Steinhauser ML, et al. Braveheart, a long noncoding RNA required for cardiovascular lineage commitment. Cell. 2013;152(3):570–83.PubMedPubMedCentralCrossRef
21.
Sun L, Goff LA, Trapnell C, Alexander R, Lo KA, Hacisuleyman E, et al. Long noncoding RNAs regulate adipogenesis. Proc Natl Acad Sci U S A. 2013;110(9):3387–92.PubMedPubMedCentralCrossRef
22.
Pang WJ, Lin LG, Xiong Y, Wei N, Wang Y, Shen QW, et al. Knockdown of PU.1 AS lncRNA inhibits adipogenesis through enhancing PU.1 mRNA translation. J Cell Biochem. 2013;114(11):2500–12.PubMedCrossRef
23.
Qiu JJ, Ye LC, Ding JX, Feng WW, Jin HY, Zhang Y, et al. Expression and clinical significance of estrogen-regulated long non-coding RNAs in estrogen receptor alpha-positive ovarian cancer progression. Oncol Rep. 2014;31(4):1613–22.PubMed
24.
Christakou C, Diamanti-Kandarakis E. Polycystic ovary syndrome—phenotypes and diagnosis. Scand J Clin Lab Invest Suppl. 2014;244:18–22.PubMedCrossRef
25.
Fux Otta C, Fiol de Cuneo M, Szafryk de Mereshian P. [Polycystic ovary syndrome: physiopathology review]. Rev Fac Cien Med Univ Nac Cordoba. 2013;70(1):27–30.PubMed
26.
Onalan G, Selam B, Baran Y, Cincik M, Onalan R, Gunduz U, et al. Serum and follicular fluid levels of soluble Fas, soluble Fas ligand and apoptosis of luteinized granulosa cells in PCOS patients undergoing IVF. Hum Reprod. 2005;20(9):2391–5.PubMedCrossRef
27.
Erickson GF, Magoffin DA, Garzo VG, Cheung AP, Chang RJ. Granulosa cells of polycystic ovaries: are they normal or abnormal? Hum Reprod. 1992;7(3):293–9.PubMed
28.
Huang X, Hao C, Shen X, Liu X, Shan Y, Zhang Y, et al. Differences in the transcriptional profiles of human cumulus cells isolated from MI and MII oocytes of patients with polycystic ovary syndrome. Reproduction. 2013;145(6):597–608.PubMedCrossRef
29.
Huang X, Hao C, Shen X, Zhang Y, Liu X. RUNX2, GPX3 and PTX3 gene expression profiling in cumulus cells are reflective oocyte/embryo competence and potentially reliable predictors of embryo developmental competence in PCOS patients. Reprod Biol Endocrinol. 2013;11:109.PubMedPubMedCentralCrossRef
30.
Orom UA, Derrien T, Beringer M, Gumireddy K, Gardini A, Bussotti G, et al. Long noncoding RNAs with enhancer-like function in human cells. Cell. 2010;143(1):46–58.PubMedPubMedCentralCrossRef
31.
Eisen MB, Spellman PT, Brown PO, Botstein D. Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci U S A. 1998;95(25):14863–8.PubMedPubMedCentralCrossRef
32.
Harrow J, Denoeud F, Frankish A, Reymond A, Chen CK, Chrast J, et al. GENCODE: producing a reference annotation for ENCODE. Genome Biol. 2006;7 Suppl 1:1–9. S4.CrossRef
33.
Guttman M, Amit I, Garber M, French C, Lin MF, Feldser D, et al. Chromatin signature reveals over a thousand highly conserved large non-coding RNAs in mammals. Nature. 2009;458(7235):223–7.PubMedPubMedCentralCrossRef
34.
Khalil AM, Guttman M, Huarte M, Garber M, Raj A, Rivea Morales D, et al. Many human large intergenic noncoding RNAs associate with chromatin-modifying complexes and affect gene expression. Proc Natl Acad Sci U S A. 2009;106(28):11667–72.PubMedPubMedCentralCrossRef
35.
Rinn JL, Kertesz M, Wang JK, Squazzo SL, Xu X, Brugmann SA, et al. Functional demarcation of active and silent chromatin domains in human HOX loci by noncoding RNAs. Cell. 2007;129(7):1311–23.PubMedPubMedCentralCrossRef
36.
Yu G, Yao W, Wang J, Ma X, Xiao W, Li H, et al. LncRNAs expression signatures of renal clear cell carcinoma revealed by microarray. PLoS One. 2012;7(8):e42377.PubMedPubMedCentralCrossRef
37.
Liao Q, Liu C, Yuan X, Kang S, Miao R, Xiao H, et al. Large-scale prediction of long non-coding RNA functions in a coding-non-coding gene co-expression network. Nucleic Acids Res. 2011;39(9):3864–78.PubMedPubMedCentralCrossRef
38.
Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498–504.PubMedPubMedCentralCrossRef
39.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001;25(4):402–8.PubMedCrossRef
40.
Wawrzik M, Spiess AN, Herrmann R, Buiting K, Horsthemke B. Expression of SNURF-SNRPN upstream transcripts and epigenetic regulatory genes during human spermatogenesis. Eur J Hum Genet. 2009;17(11):1463–70.PubMedPubMedCentralCrossRef
41.
Sun M, Kraus WL. Minireview: long noncoding RNAs: new “links” between gene expression and cellular outcomes in endocrinology. Mol Endocrinol. 2013;27(9):1390–402.PubMedPubMedCentralCrossRef
42.
Sun PR, Jia SZ, Lin H, Leng JH, Lang JH. Genome-wide profiling of long noncoding ribonucleic acid expression patterns in ovarian endometriosis by microarray. Fertil Steril. 2014;101(4):1038–46. e7.PubMedCrossRef
43.
Chen ZJ, Zhao H, He L, Shi Y, Qin Y, Li Z, et al. Genome-wide association study identifies susceptibility loci for polycystic ovary syndrome on chromosome 2p16.3, 2p21 and 9q33.3. Nat Genet. 2011;43(1):55–9.PubMedCrossRef
44.
Xu B, Gerin I, Miao H, Vu-Phan D, Johnson CN, Xu R, et al. Multiple roles for the non-coding RNA SRA in regulation of adipogenesis and insulin sensitivity. PLoS One. 2010;5(12):e14199.PubMedPubMedCentralCrossRef
45.
Kino T, Hurt DE, Ichijo T, Nader N, Chrousos GP. Noncoding RNA gas5 is a growth arrest- and starvation-associated repressor of the glucocorticoid receptor. Sci Signal. 2010;3(107):ra8.PubMedPubMedCentral
46.
Takayama K, Horie-Inoue K, Katayama S, Suzuki T, Tsutsumi S, Ikeda K, et al. Androgen-responsive long noncoding RNA CTBP1-AS promotes prostate cancer. EMBO J. 2013;32(12):1665–80.PubMedPubMedCentralCrossRef
47.
Wang L, Rao F, Zhang K, Mahata M, Rodriguez-Flores JL, Fung MM, et al. Neuropeptide Y(1) Receptor NPY1R discovery of naturally occurring human genetic variants governing gene expression in cella as well as pleiotropic effects on autonomic activity and blood pressure in vivo. J Am Coll Cardiol. 2009;54(10):944–54.PubMedPubMedCentralCrossRef
48.
Elbers CC, Onland-Moret NC, Franke L, Niehoff AG, van der Schouw YT, Wijmenga C. A strategy to search for common obesity and type 2 diabetes genes. Trends Endocrinol Metab. 2007;18(1):19–26.PubMedCrossRef
49.
Gericke MT, Schroder T, Kosacka J, Nowicki M, Kloting N, Spanel-Borowski K. Neuropeptide Y impairs insulin-stimulated translocation of glucose transporter 4 in 3T3-L1 adipocytes through the Y1 receptor. Mol Cell Endocrinol. 2012;348(1):27–32.PubMed
50.
Pietrement C, Sun-Wada GH, Silva ND, McKee M, Marshansky V, Brown D, et al. Distinct expression patterns of different subunit isoforms of the V-ATPase in the rat epididymis. Biol Reprod. 2006;74(1):185–94.PubMedCrossRef
51.
Van Blerkom J. The influence of intrinsic and extrinsic factors on the developmental potential and chromosomal normality of the human oocyte. J Soc Gynecol Investig. 1996;3(1):3–11.PubMedCrossRef
52.
Nagano T, Mitchell JA, Sanz LA, Pauler FM, Ferguson-Smith AC, Feil R, et al. The Air noncoding RNA epigenetically silences transcription by targeting G9a to chromatin. Science. 2008;322(5908):1717–20.PubMedCrossRef
Metadata
Title
Aberrant expression of long noncoding RNAs in cumulus cells isolated from PCOS patients
Authors
Xin Huang
Cuifang Hao
Hongchu Bao
Meimei Wang
Huangguan Dai
Publication date
01-01-2016
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 1/2016
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-015-0630-z

Other articles of this Issue 1/2016

Journal of Assisted Reproduction and Genetics 1/2016 Go to the issue

Genetics

Association of single-nucleotide polymorphisms rs2197076 and rs2241883 of FABP1 gene with polycystic ovary syndrome

Opinion

Is the wrong question being asked in infertility research?

Assisted Reproduction Technologies

Independent factors influencing large-for-gestation birth weight in singletons born after in vitro fertilization

Assisted Reproduction Technologies

Does reducing gamete co-incubation time improve clinical outcomes: a retrospective study

Reproductive Physiology and Disease

The anti-Müllerian hormone (AMH) acts as a gatekeeper of ovarian steroidogenesis inhibiting the granulosa cell response to both FSH and LH

Oocyte Biology

ART results with frozen oocytes: data from the Italian ART registry (2005–2013)