期刊论文详细信息
Reproductive Biology and Endocrinology
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
Xiaoyan Liu2  Yuhua Zhang2  Xiaofang Shen2  Cuifang Hao2  Xin Huang1 
[1] CapitalBio Corporation Center (Biochip National Engineering Research Center), Yantai Branch Center, 20 Yuhuangding Road East, Yantai, Shandong, China;Reproductive Medicine Centre, Affiliated Hospital of Qingdao Medical University, Yuhuangding Hospital of Yantai, 20 Yuhuangding Road East, Yantai, Shandong, China
关键词: PCOS;    Oocyte maturation;    Human cumulus cells;    Gene expression;    Fertilization;    Embryo development;   
Others  :  809846
DOI  :  10.1186/1477-7827-11-109
 received in 2013-08-28, accepted in 2013-11-23,  发布年份 2013
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【 摘 要 】

Background

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. The developmental competence of oocytes and embryos in PCOS patients is reduced to a certain extent (comparing to non-PCOS patients, the high quality embryo rate was decreased by 16% from the data of our centre) during the in vitro fertilization (IVF) process. Cross-talk between the oocyte and cumulus cells is critical for oocyte maturation and embryo competence. In this study, we have evaluated the transcription of specific genes in cumulus cells harvested from pre-ovulatory follicles of PCOS patients before IVF, according to individual oocyte nuclear maturity and developmental competence. Seven genes (RUNX2, PSAT1, ADAMTS9, CXCL1, CXCL2, CXCL3, and ITGB5) were targeted from our previous cDNA microarray data which isolated genes related to oocyte nuclear maturation in PCOS patients. Two additional genes which had been found to be associated with oocyte maturation or embryo quality in non-PCOS patients (GPX3 and PTX3) were also studied.

Methods

The mRNA expression levels of cumulus cells were detected by qRT- PCR.

Results

Consistent with our previous cDNA microarray data, with the exception of GPX3 and PTX3, the selected 7 genes were related to oocyte nuclear maturation in PCOS patients. Noticeably, the expression level of RUNX2 was lower in cumulus cells derived from oocytes that could develop into blastocysts than the level of expression from oocytes that could not. The PTX3 expression level was significantly lower in cumulus cells from oocytes with two normal pronuclei than that from oocytes that formed >2 pronuclei (MPN) after fertilization. GPX3 mRNA levels were decreased in cumulus cells isolated from oocytes that developed into blastocysts with high potential development competence.

Conclusions

Several cumulus cell genes were associated with oocyte maturation, fertilization and embryo quality in PCOS patients. RUNX2 and GPX3 are candidate genetic markers in the monitoring of embryo quality for PCOS patients, whereas PTX3 mainly played a role in fertilization process. Together with morphological evaluation, cumulus cells genes may serve as biomarkers of oocyte and embryo selection during the IVF process for PCOS patients and may advance our understanding of PCOS.

【 授权许可】

   
2013 Huang et al.; licensee BioMed Central Ltd.

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【 参考文献 】
  • [1]Franks S: Polycystic ovary syndrome. N Engl J Med 1995, 333:853-861.
  • [2]Legro RS: Polycystic ovary syndrome: the new millenium. Mol Cell Endocrinol 2001, 184:87-93.
  • [3]Ehrmann DA: Polycystic ovary syndrome. N Engl J Med 2005, 352:1223-1236.
  • [4]Wood JR, Nelson VL, Ho C, Jansen E, Wang CY, Urbanek M, McAllister JM, Mosselman S, Strauss JF 3rd: The molecular phenotype of polycystic ovary syndrome (PCOS) theca cells and new candidate PCOS genes defined by microarray analysis. J Biol Chem 2003, 278:26380-26390.
  • [5]Wood JR, Dumesic DA, Abbott DH, Strauss JF 3rd: Molecular abnormalities in oocytes from women with polycystic ovary syndrome revealed by microarray analysis. J Clin Endocrinol Metab 2007, 92:705-713.
  • [6]Kwon H, Choi DH, Bae JH, Kim JH, Kim YS: mRNA expression pattern of insulin-like growth factor components of granulosa cells and cumulus cells in women with and without polycystic ovary syndrome according to oocyte maturity. Fertil Steril 2010, 94:2417-2420.
  • [7]Liu Y, Jiang H, He LY, Huang WJ, He XY, Xing FQ: Abnormal expression of uncoupling protein-2 correlates with CYP11A1 expression in polycystic ovary syndrome. Reprod Fertil Dev 2011, 23:520-526.
  • [8]Glueck CJ, Wang P, Goldenberg N, Sieve-Smith L: Pregnancy outcomes among women with polycystic ovary syndrome treated with metformin. Hum Reprod 2002, 17:2858-2864.
  • [9]Arredondo F, Noble LS: Endocrinology of recurrent pregnancy loss. Semin Reprod Med 2006, 24:33-39.
  • [10]Ouandaogo ZG, Haouzi D, Assou S, Dechaud H, Kadoch IJ, De Vos J, Hamamah S: Human cumulus cells molecular signature in relation to oocyte nuclear maturity stage. PLoS One 2011, 6:e27179.
  • [11]Ouandaogo ZG, Frydman N, Hesters L, Assou S, Haouzi D, Dechaud H, Frydman R, Hamamah S: Differences in transcriptomic profiles of human cumulus cells isolated from oocytes at GV, MI and MII stages after in vivo and in vitro oocyte maturation. Hum Reprod 2012, 27:2438-2447.
  • [12]Gasca S, Pellestor F, Assou S, Loup V, Anahory T, Dechaud H, De Vos J, Hamamah S: Identifying new human oocyte marker genes: a microarray approach. Reprod Biomed Online 2007, 14:175-183.
  • [13]Huang Z, Wells D: The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol Hum Reprod 2010, 16:715-725.
  • [14]Assou S, Haouzi D, De Vos J, Hamamah S: Human cumulus cells as biomarkers for embryo and pregnancy outcomes. Mol Hum Reprod 2010, 16:531-538.
  • [15]Cross PC, Brinster RL: In vitro development of mouse oocytes. Biol Reprod 1970, 3:298-307.
  • [16]Goud PT, Goud AP, Qian C, Laverge H, Van der Elst J, De Sutter P, Dhont M: In-vitro maturation of human germinal vesicle stage oocytes: role of cumulus cells and epidermal growth factor in the culture medium. Hum Reprod 1998, 13:1638-1644.
  • [17]McKenzie LJ, Pangas SA, Carson SA, Kovanci E, Cisneros P, Buster JE, Amato P, Matzuk MM: Human cumulus granulosa cell gene expression: a predictor of fertilization and embryo selection in women undergoing IVF. Hum Reprod 2004, 19:2869-2874.
  • [18]Zhang X, Jafari N, Barnes RB, Confino E, Milad M, Kazer RR: Studies of gene expression in human cumulus cells indicate pentraxin 3 as a possible marker for oocyte quality. Fertil Steril 2005, 83(Suppl 1):1169-1179.
  • [19]Feuerstein P, Cadoret V, Dalbies-Tran R, Guerif F, Bidault R, Royere D: Gene expression in human cumulus cells: one approach to oocyte competence. Hum Reprod 2007, 22:3069-3077.
  • [20]Assou S, Haouzi D, Mahmoud K, Aouacheria A, Guillemin Y, Pantesco V, Reme T, Dechaud H, De Vos J, Hamamah S: A non-invasive test for assessing embryo potential by gene expression profiles of human cumulus cells: a proof of concept study. Mol Hum Reprod 2008, 14:711-719.
  • [21]Hamel M, Dufort I, Robert C, Gravel C, Leveille MC, Leader A, Sirard MA: Identification of differentially expressed markers in human follicular cells associated with competent oocytes. Hum Reprod 2008, 23:1118-1127.
  • [22]van Montfoort AP, Geraedts JP, Dumoulin JC, Stassen AP, Evers JL, Ayoubi TA: Differential gene expression in cumulus cells as a prognostic indicator of embryo viability: a microarray analysis. Mol Hum Reprod 2008, 14:157-168.
  • [23]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, 15:89-103.
  • [24]Adriaenssens T, Wathlet S, Segers I, Verheyen G, De Vos A, Van der Elst J, Coucke W, Devroey P, Smitz J: Cumulus cell gene expression is associated with oocyte developmental quality and influenced by patient and treatment characteristics. Hum Reprod 2010, 25:1259-1270.
  • [25]Huang X, Hao C, Shen X, Liu X, Shan Y, Zhang Y, Chen L: Differences in the transcriptional profiles of human cumulus cells isolated from MI and MII oocytes of patients with polycystic ovary syndrome. Reproduction 2013, 145:597-608.
  • [26]Behr B, Pool TB, Milki AA, Moore D, Gebhardt J, Dasig D: Preliminary clinical experience with human blastocyst development in vitro without co-culture. Hum Reprod 1999, 14:454-457.
  • [27]Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T: The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol 2006, 7:3. BioMed Central Full Text
  • [28]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, 25:402-408.
  • [29]Dumesic DA, Abbott DH: Implications of polycystic ovary syndrome on oocyte development. Semin Reprod Med 2008, 26:53-61.
  • [30]Ludwig M, Finas DF, Al-Hasani S, Diedrich K, Ortmann O: Oocyte quality and treatment outcome in intracytoplasmic sperm injection cycles of polycystic ovarian syndrome patients. Hum Reprod 1999, 14:354-358.
  • [31]Baek JY, Jun DY, Taub D, Kim YH: Characterization of human phosphoserine aminotransferase involved in the phosphorylated pathway of L-serine biosynthesis. Biochem J 2003, 373:191-200.
  • [32]Vie N, Copois V, Bascoul-Mollevi C, Denis V, Bec N, Robert B, Fraslon C, Conseiller E, Molina F, Larroque C, et al.: Overexpression of phosphoserine aminotransferase PSAT1 stimulates cell growth and increases chemoresistance of colon cancer cells. Mol Cancer 2008, 7:14. BioMed Central Full Text
  • [33]Jungers KA, Le Goff C, Somerville RP, Apte SS: Adamts9 is widely expressed during mouse embryo development. Gene Expr Patterns 2005, 5:609-617.
  • [34]Irving-Rodgers HF, Rodgers RJ: Extracellular matrix in ovarian follicular development and disease. Cell Tissue Res 2005, 322:89-98.
  • [35]Bromfield JJ, Sheldon IM: Lipopolysaccharide initiates inflammation in bovine granulosa cells via the TLR4 pathway and perturbs oocyte meiotic progression in vitro. Endocrinology 2011, 152:5029-5040.
  • [36]Hannan NJ, Salamonsen LA: Role of chemokines in the endometrium and in embryo implantation. Curr Opin Obstet Gynecol 2007, 19:266-272.
  • [37]Dowsland MH, Harvey JR, Lennard TW, Kirby JA, Ali S: Chemokines and breast cancer: a gateway to revolutionary targeted cancer treatments? Curr Med Chem 2003, 10:579-592.
  • [38]Aplin JD, Spanswick C, Behzad F, Kimber SJ, Vicovac L: Integrins beta 5, beta 3 and alpha v are apically distributed in endometrial epithelium. Mol Hum Reprod 1996, 2:527-534.
  • [39]Hannan NJ, Salamonsen LA: CX3CL1 and CCL14 regulate extracellular matrix and adhesion molecules in the trophoblast: potential roles in human embryo implantation. Biol Reprod 2008, 79:58-65.
  • [40]Elvin JA, Clark AT, Wang P, Wolfman NM, Matzuk MM: Paracrine actions of growth differentiation factor-9 in the mammalian ovary. Mol Endocrinol 1999, 13:1035-1048.
  • [41]Chang H, Brown CW, Matzuk MM: Genetic analysis of the mammalian transforming growth factor-beta superfamily. Endocr Rev 2002, 23:787-823.
  • [42]Vanderhyden BC, Macdonald EA, Nagyova E, Dhawan A: Evaluation of members of the TGFbeta superfamily as candidates for the oocyte factors that control mouse cumulus expansion and steroidogenesis. Reprod Suppl 2003, 61:55-70.
  • [43]Salustri A, Garlanda C, Hirsch E, De Acetis M, Maccagno A, Bottazzi B, Doni A, Bastone A, Mantovani G, Beck Peccoz P, et al.: PTX3 plays a key role in the organization of the cumulus oophorus extracellular matrix and in in vivo fertilization. Development 2004, 131:1577-1586.
  • [44]Varani S, Elvin JA, Yan C, DeMayo J, DeMayo FJ, Horton HF, Byrne MC, Matzuk MM: Knockout of pentraxin 3, a downstream target of growth differentiation factor-9, causes female subfertility. Mol Endocrinol 2002, 16:1154-1167.
  • [45]Cillo F, Brevini TA, Antonini S, Paffoni A, Ragni G, Gandolfi F: Association between human oocyte developmental competence and expression levels of some cumulus genes. Reproduction 2007, 134:645-650.
  • [46]Pangas SA, Li X, Robertson EJ, Matzuk MM: Premature luteinization and cumulus cell defects in ovarian-specific Smad4 knockout mice. Mol Endocrinol 2006, 20:1406-1422.
  • [47]Park ES, Lind AK, Dahm-Kahler P, Brannstrom M, Carletti MZ, Christenson LK, Curry TE Jr, Jo M: RUNX2 transcription factor regulates gene expression in luteinizing granulosa cells of rat ovaries. Mol Endocrinol 2010, 24:846-858.
  • [48]Fan HY, Liu Z, Johnson PF, Richards JS: CCAAT/enhancer-binding proteins (C/EBP)-alpha and -beta are essential for ovulation, luteinization, and the expression of key target genes. Mol Endocrinol 2011, 25:253-268.
  • [49]Papamentzelopoulou M, Mavrogianni D, Dinopoulou V, Theofanakis H, Malamas F, Marinopoulos S, Bletsa R, Anagnostou E, Kallianidis K, Loutradis D: Detection of RUNX2 gene expression in cumulus cells in women undergoing controlled ovarian stimulation. Reprod Biol Endocrinol 2012, 10:99. BioMed Central Full Text
  • [50]Schmidt J, Weijdegard B, Mikkelsen AL, Lindenberg S, Nilsson L, Brannstrom M: Differential expression of inflammation-related genes in the ovarian stroma and granulosa cells of PCOS women. Mol Hum Reprod 2013. Aug 20. [Epub ahead of print]
  • [51]Wathlet S, Adriaenssens T, Segers I, Verheyen G, Van Landuyt L, Coucke W, Devroey P, Smitz J: Pregnancy prediction in single embryo transfer cycles after ICSI using QPCR: validation in oocytes from the same cohort. PLoS One 2013, 8:e54226.
  • [52]Bierl C, Voetsch B, Jin RC, Handy DE, Loscalzo J: Determinants of human plasma glutathione peroxidase (GPx-3) expression. J Biol Chem 2004, 279:26839-26845.
  • [53]Buttke TM, Sandstrom PA: Oxidative stress as a mediator of apoptosis. Immunol Today 1994, 15:7-10.
  • [54]Tatemoto H, Sakurai N, Muto N: Protection of porcine oocytes against apoptotic cell death caused by oxidative stress during In vitro maturation: role of cumulus cells. Biol Reprod 2000, 63:805-810.
  • [55]Pasqualotto EB, Agarwal A, Sharma RK, Izzo VM, Pinotti JA, Joshi NJ, Rose BI: Effect of oxidative stress in follicular fluid on the outcome of assisted reproductive procedures. Fertil Steril 2004, 81:973-976.
  • [56]Das S, Chattopadhyay R, Ghosh S, Goswami SK, Chakravarty BN, Chaudhury K: Reactive oxygen species level in follicular fluid–embryo quality marker in IVF? Hum Reprod 2006, 21:2403-2407.
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