Genome‑wide DNA methylation analysis of uterosacral ligaments in women with pelvic organ prolapse

Zhang,Lifang; Zheng,Ping; Duan,Aihong; Hao,Yan; Lu,Chang; Lu,Dan

doi:10.3892/mmr.2018.9656

Genome‑wide DNA methylation analysis of uterosacral ligaments in women with pelvic organ prolapse

Authors:
- Lifang Zhang
- Ping Zheng
- Aihong Duan
- Yan Hao
- Chang Lu
- Dan Lu
View Affiliations

Affiliations: Department of Gynecology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, P.R. China
Published online on: November 13, 2018 https://doi.org/10.3892/mmr.2018.9656
Pages: 391-399
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Pelvic organ prolapse (POP) is an increasingly serious health problem that impairs quality of life and is caused by multiple additive genetic and environmental factors. As the uterosacral ligaments (ULs) provide primary support for the pelvic organs, it was hypothesized that disruption of these ligaments (as a result of aberrant methylation) may lead to a loss of support and eventually contribute to POP. In the present study, whether there are any aberrant methylations in the ULs of patients with POP compared to those of controls was investigated. Genomic DNA was isolated from the ULs of five women with POP and four women without POP, as controls, undergoing hysterectomy for benign conditions. An Illumina Infinium Methylation EPICBeadChips Infinium Human Methylation 850 K bead array was used to investigate the total methylation in the ULs. There were 3,723 differentially methylated CpG sites (Δβ<0.14; P<0.05), including 3,576 hypermethylation and 147 hypomethylation sites in the ULs of patients with POP compared with the normal controls. There were more hypermethylated CpG sites, but a high ratio of hypomethylation between CpG islands and the N‑shelf; in the gene structure, there was more hypermethylation than hypomethylation in TSS1500 and the 5' untranslated region. Gene ontology analysis demonstrated that these differentially methylated genes were associated with ‘cell morphogenesis’, ‘extracellular matrix’, ‘cell junction’, ‘protein binding’ and ‘guanosine triphosphatase activity’. Several significant pathways were identified, including ‘focal adhesion’ and ‘extracellular matrix‑receptor interaction pathway’. This study provides evidence that there are differences in genome‑wide DNA methylation between ULs in menopausal women with and without POP, and that epigenetic mechanisms may partly contribute to POP pathogenesis.

View Figures

View References

1	DeLancey JO: The hidden epidemic of pelvic floor dysfunction: Achievable goals for improved prevention and treatment. Am J Obstet Gynecol. 192:1488–1495. 2005. View Article : Google Scholar : PubMed/NCBI
2	Nygaard I, Barber MD, Burgio KL, Kenton K, Meikle S, Schaffer J, Spino C, Whitehead WE, Wu J and Brody DJ: Pelvic Floor Disorders Network: Prevalence of symptomatic pelvic floor disorders in US women. JAMA. 300:1311–1316. 2008. View Article : Google Scholar : PubMed/NCBI
3	Whitcomb EL, Rortveit G, Brown JS, Creasman JM, Thom DH, Van Den Eeden SK and Subak LL: Racial differences in pelvic organ prolapse. Obstet Gynecol. 114:1271–1277. 2009. View Article : Google Scholar : PubMed/NCBI
4	Rortveit G, Brown JS, Thom DH, Van Den Eeden SK, Creasman JM and Subak LL: Symptomatic pelvic organ prolapse: Prevalence and risk factors in a population-based, racially diverse cohort. Obstet Gynecol. 109:1396–1403. 2007. View Article : Google Scholar : PubMed/NCBI
5	Hendrix SL, Clark A, Nygaard I, Aragaki A, Barnabei V and McTiernan A: Pelvic organ prolapse in the Women's Health Initiative: Gravity and gravidity. Am J Obstet Gynecol. 186:1160–1166. 2002. View Article : Google Scholar : PubMed/NCBI
6	Glazener C, Elders A, Macarthur C, Lancashire RJ, Herbison P, Hagen S, Dean N, Bain C, Toozs-Hobson P, Richardson K, et al: Childbirth and prolapse: Long-term associations with the symptoms and objective measurement of pelvic organ prolapse. BJOG. 120:161–168. 2013. View Article : Google Scholar : PubMed/NCBI
7	Bradley CS, Zimmerman MB, Qi Y and Nygaard IE: Natural history of pelvic organ prolapse in postmenopausal women. Obstet Gynecol. 109:848–854. 2007. View Article : Google Scholar : PubMed/NCBI
8	Kudish BI, Iglesia CB, Sokol RJ, Cochrane B, Richter HE, Larson J, Hendrix SL and Howard BV: Effect of weight change on natural history of pelvic organ prolapse. Obstet Gynecol. 113:81–88. 2009. View Article : Google Scholar : PubMed/NCBI
9	Xie R, Xu Y, Fan S and Song Y: Identification of differentially expressed genes in pelvic organ prolapse by RNA-Seq. Med Sci Monit. 22:4218–4225. 2016. View Article : Google Scholar : PubMed/NCBI
10	Ak H, Zeybek B, Atay S, Askar N, Akdemir A and Aydin HH: Microarray gene expression analysis of uterosacral ligaments in uterine prolapse. Clin Biochem. 49:1238–1242. 2016. View Article : Google Scholar : PubMed/NCBI
11	Allen-Brady K, Cannon-Albright L, Farnham JM, Teerlink C, Vierhout ME, van Kempen LC, Kluivers KB and Norton PA: Identification of six loci associated with pelvic organ prolapse using genome-wide association analysis. Obstet Gynecol. 118:1345–1353. 2011. View Article : Google Scholar : PubMed/NCBI
12	Allen-Brady K, Cannon-Albright LA, Farnham JM and Norton PA: Evidence for pelvic organ prolapse predisposition genes on chromosomes 10 and 17. Am J Obstet Gynecol. 212:771.e1–e7. 2015. View Article : Google Scholar
13	Rao S, Lang J, Zhu L and Chen J: Exome sequencing identifies a novel gene, WNK1, for susceptibility to pelvic organ prolapse (POP). PLoS One. 10:e01194822015. View Article : Google Scholar : PubMed/NCBI
14	Sui X, Zhu J, Zhou J, Wang X, Li D, Han W, Fang Y and Pan H: Epigenetic modifications as regulatory elements of autophagy in cancer. Cancer Lett. 360:106–113. 2015. View Article : Google Scholar : PubMed/NCBI
15	Klutke J, Stanczyk FZ, Ji Q, Campeau JD and Klutke CG: Suppression of lysyl oxidase gene expression by methylation in pelvic organ prolapse. Int Urogynecol J. 21:869–872. 2010. View Article : Google Scholar : PubMed/NCBI
16	Aryee MJ, Jaffe AE, Corrada-Bravo H, Ladd-Acosta C, Feinberg AP, Hansen KD and Irizarry RA: Minfi: A flexible and comprehensive Bioconductor package for the analysis of Infinium DNA methylation microarrays. Bioinformatics. 30:1363–1369. 2014. View Article : Google Scholar : PubMed/NCBI
17	Bird AP and Wolffe AP: Methylation-induced repression-belts, braces, and chromatin. Cell. 99:451–454. 1999. View Article : Google Scholar : PubMed/NCBI
18	Christensen BC, Houseman EA, Marsit CJ, Zheng S, Wrensch MR, Wiemels JL, Nelson HH, Karagas MR, Padbury JF, Bueno R, et al: Aging and environmental exposures alter tissue-specific DNA methylation dependent upon CpG island context. PLoS Genet. 5:e10006022009. View Article : Google Scholar : PubMed/NCBI
19	Rang FJ and Boonstra J: Causes and consequences of age-related changes in DNA methylation: A Role for ROS? Biology (Basel). 3:403–425. 2014.PubMed/NCBI
20	Borazjani A, Kow N, Harris S, Ridgeway B and Damaser MS: Transcriptional regulation of connective tissue metabolism genes in women with pelvic organ prolapse. Female Pelvic Med Reconstr Surg. 23:44–52. 2017. View Article : Google Scholar : PubMed/NCBI
21	Sun MJ, Cheng YS, Sun R, Cheng WL and Liu CS: Changes in mitochondrial DNA copy number and extracellular matrix (ECM) proteins in the uterosacral ligaments of premenopausal women with pelvic organ prolapse. Taiwan J Obstet Gynecol. 55:9–15. 2016. View Article : Google Scholar : PubMed/NCBI
22	Connell KA, Guess MK, Chen H, Andikyan V, Bercik R and Taylor HS: HOXA11 is critical for development and maintenance of uterosacral ligaments and deficient in pelvic prolapse. J Clin Invest. 118:1050–1055. 2008.PubMed/NCBI
23	Alarab M, Bortolini MA, Drutz H, Lye S and Shynlova O: LOX family enzymes expression in vaginal tissue of premenopausal women with severe pelvic organ prolapse. Int Urogynecol J. 21:1397–1404. 2010. View Article : Google Scholar : PubMed/NCBI
24	Kobak W, Lu J, Hardart A, Zhang C, Stanczyk FZ and Felix JC: Expression of lysyl oxidase and transforming growth factor beta2 in women with severe pelvic organ prolapse. J Reprod Med. 50:827–831. 2005.PubMed/NCBI
25	Klutke J, Ji Q, Campeau J, Starcher B, Felix JC, Stanczyk FZ and Klutke C: Decreased endopelvic fascia elastin content in uterine prolapse. Acta Obstet Gynecol Scand. 87:111–115. 2008. View Article : Google Scholar : PubMed/NCBI
26	Wang X, Li Y, Chen J, Guo X, Guan H and Li C: Differential expression profiling of matrix metalloproteinases and tissue inhibitors of metalloproteinases in females with or without pelvic organ prolapse. Mol Med Rep. 10:2004–2008. 2014. View Article : Google Scholar : PubMed/NCBI
27	Alarab M, Kufaishi H, Lye S, Drutz H and Shynlova O: Expression of extracellular matrix-remodeling proteins is altered in vaginal tissue of premenopausal women with severe pelvic organ prolapse. Reprod Sci. 21:704–715. 2014. View Article : Google Scholar : PubMed/NCBI
28	Usta A, Guzin K, Kanter M, Ozgul M and Usta CS: Expression of matrix metalloproteinase-1 in round ligament and uterosacral ligament tissue from women with pelvic organ prolapse. J Mol Histol. 45:275–281. 2014. View Article : Google Scholar : PubMed/NCBI
29	Yilmaz N, Ozaksit G, Terzi YK, Yilmaz S, Budak B, Aksakal O and Sahin FI: HOXA11 and MMP2 gene expression in uterosacral ligaments of women with pelvic organ prolapse. J Turk Ger Gynecol Assoc. 15:104–108. 2014. View Article : Google Scholar : PubMed/NCBI
30	Liang CC, Huang HY and Chang SD: Gene expression and immunoreactivity of elastolytic enzymes in the uterosacral ligaments from women with uterine prolapse. Reprod Sci. 19:354–359. 2012. View Article : Google Scholar : PubMed/NCBI
31	Ma Y, Guess M, Datar A, Hennessey A, Cardenas I, Johnson J and Connell KA: Knockdown of Hoxa11 in vivo in the uterosacral ligament and uterus of mice results in altered collagen and matrix metalloproteinase activity. Biol Reprod. 86:1002012. View Article : Google Scholar : PubMed/NCBI
32	Connell KA, Guess MK, Chen HW, Lynch T, Bercik R and Taylor HS: HOXA11 promotes fibroblast proliferation and regulates p53 in uterosacral ligaments. Reprod Sci. 16:694–700. 2009. View Article : Google Scholar : PubMed/NCBI
33	Kufaishi H, Alarab M, Drutz H, Lye S and Shynlova O: Comparative characterization of vaginal cells derived from premenopausal women with and without severe pelvic organ prolapse. Reprod Sci. 23:931–943. 2016. View Article : Google Scholar : PubMed/NCBI
34	Kufaishi H, Alarab M, Drutz H, Lye S and Shynlova O: Static mechanical loading influences the expression of extracellular matrix and cell adhesion proteins in vaginal cells derived from premenopausal women with severe pelvic organ prolapse. Reprod Sci. 23:978–992. 2016. View Article : Google Scholar : PubMed/NCBI
35	Li BS, Guo WJ, Hong L, Liu YD, Liu C, Hong SS, Wu DB and Min J: Role of mechanical strain-activated PI3K/Akt signaling pathway in pelvic organ prolapse. Mol Med Rep. 14:243–253. 2016. View Article : Google Scholar : PubMed/NCBI
36	Chen X, Kong X, Liu D, Gao P, Zhang Y, Li P and Liu M: In vitro differentiation of endometrial regenerative cells into smooth muscle cells: Alpha potential approach for the management of pelvic organ prolapse. Int J Mol Med. 38:95–104. 2016. View Article : Google Scholar : PubMed/NCBI
37	Giancotti FG and Ruoslahti E: Integrin signaling. Science. 285:1028–1032. 1999. View Article : Google Scholar : PubMed/NCBI
38	Rhee SH, Zhang P, Hunter K, Mama ST, Caraballo R, Holzberg AS, Seftel RH, Seftel AD, Echols KT and DiSanto ME: Pelvic organ prolapse is associated with alteration of sphingosine-1-phosphate/Rho-kinase signalling pathway in human vaginal wall. J Obstet Gynaecol. 35:726–732. 2015. View Article : Google Scholar : PubMed/NCBI
39	Pavlidis P, Li Q and Noble WS: The effect of replication on gene expression microarray experiments. Bioinformatics. 19:1620–1627. 2003. View Article : Google Scholar : PubMed/NCBI