The association between rapsn methylation in peripheral blood and breast cancer in the chinese population

ABSTRACT DNA methylation in peripheral blood is associated with breast cancer (BC) but has mainly been studied in Caucasian populations. We investigated the association between blood-based

methylation of receptor-associated protein of the synapse (_RAPSN_) and BC in Chinese population. The methylation levels of 12 _RAPSN_ CpG sites were quantitatively evaluated by mass

spectrometry in two case–control studies with 283 sporadic BC cases and 331 controls totally. The association was analyzed by logistic regression adjusted for covariants. The _RAPSN_

methylation levels in patients with variant clinical characteristics were investigated by non-parametric tests. We found a significant association between BC and altered _RAPSN_ methylation

in blood in women at premenopausal and perimenopausal (age < 50 years old), but not in the elder women. This was approved by two independent case–control studies as well as by combining

the subjects of the two studies (taken all subjects together, age < 50 years old, per 5% of methylation, odds ratio (OR) range from 1.17 to 1.30 for two CpG sites; OR = 0.75 for one CpG

site; all _p_ values < 0.02). This age-related _RAPSN_ methylation was further modified by human epidermal growth factor receptor 2 (HER2) status (age < 50 years old, HER2 negative,

per 5% of methylation, OR range from 1.27 to 1.48 for two CpG sites; OR = 0.76 for one CpG site; all _p_ values < 0.02). We elucidated an association between BC and blood-based _RAPSN_

methylation influenced by age and the status of HER2 in Chinese population. Access through your institution Buy or subscribe This is a preview of subscription content, access via your

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access 16 July 2020 REFERENCES * Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for

36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–24. PubMed  Google Scholar  * Momenimovahed Z, Salehiniya H. Epidemiological characteristics of 577and risk factors for breast

cancer in the world. Breast Cancer. 2019;11:151–64. PubMed  PubMed Central  Google Scholar  * Rivera-Franco MM, Leon-Rodriguez E. Delays in breast cancer detection and treatment in

developing countries. Breast Cancer. 2018;12:1178223417752677. PubMed  PubMed Central  Google Scholar  * Jafari SH, Saadatpour Z, Salmaninejad A, Momeni F, Mokhtari M, Nahand JS, et al.

Breast cancer diagnosis: imaging techniques and biochemical markers. J Cell Physiol. 2018;233:5200–13. Article  CAS  PubMed  Google Scholar  * Baylin SB, Jones PA. Epigenetic Determinants of

Cancer. Cold Spring Harb Perspect Biol. 2016;8. https://doi.org/10.1101/cshperspect.a019505. * Jankowska AM, Millward CL, Caldwell CW. The potential of DNA modifications as biomarkers and

therapeutic targets in oncology. Expert Rev Mol Diagn. 2015;15:1325–37. Article  CAS  PubMed  Google Scholar  * Craig JM, Bickmore WA. The distribution of CpG islands in mammalian

chromosomes. Nat Genet. 1994;7:376–82. Article  CAS  PubMed  Google Scholar  * Watanabe Y, Maekawa M. Methylation of DNA in cancer. Adv Clin Chem. 2010;52:145–67. Article  CAS  PubMed 

Google Scholar  * Ehrlich M. Cancer-linked DNA hypomethylation and its relationship to hypermethylation. Curr Top Microbiol Immunol. 2006;310:251–74. CAS  PubMed  Google Scholar  *

Subramaniam D, Thombre R, Dhar A, Anant S. DNA methyltransferases: a novel target for prevention and therapy. Front Oncol. 2014;4:80. Article  PubMed  PubMed Central  Google Scholar  *

Gyorffy B, Bottai G, Fleischer T, Munkacsy G, Budczies J, Paladini L, et al. Aberrant DNA methylation impacts gene expression and prognosis in breast cancer subtypes. Int J Cancer.

2016;138:87–97. Article  CAS  PubMed  Google Scholar  * Crowley E, Di Nicolantonio F, Loupakis F, Bardelli A. Liquid biopsy: monitoring cancer-genetics in the blood. Nat Rev Clin Oncol.

2013;10:472–84. Article  CAS  PubMed  Google Scholar  * Tang Q, Cheng J, Cao X, Surowy H, Burwinkel B. Blood-based DNA methylation as biomarker for breast cancer: a systematic review. Clin

Epigenetics. 2016;8:115. Article  PubMed  PubMed Central  CAS  Google Scholar  * Brennan K, Garcia-Closas M, Orr N, Fletcher O, Jones M, Ashworth A, et al. Intragenic ATM methylation in

peripheral blood DNA as a biomarker of breast cancer risk. Cancer Res. 2012;72:2304–13. Article  CAS  PubMed  Google Scholar  * Hoffman AE, Yi CH, Zheng T, Stevens RG, Leaderer D, Zhang Y,

et al. CLOCK in breast tumorigenesis: genetic, epigenetic, and transcriptional profiling analyses. Cancer Res. 2010;70:1459–68. Article  CAS  PubMed  PubMed Central  Google Scholar  * Tang

Q, Holland-Letz T, Slynko A, Cuk K, Marme F, Schott S, et al. DNA methylation array analysis identifies breast cancer associated RPTOR, MGRN1 and RAPSN hypomethylation in peripheral blood

DNA. Oncotarget. 2016;7:64191–202. Article  PubMed  PubMed Central  Google Scholar  * Yang R, Stocker S, Schott S, Heil J, Marme F, Cuk K, et al. The association between breast cancer and

S100P methylation in peripheral blood by multicenter case-control studies. Carcinogenesis. 2017;38:312–20. Article  CAS  PubMed  Google Scholar  * Yin Q, Yang X, Li L, Xu T, Zhou W, Gu W, et

al. The association between breast cancer and blood-based methylation of S100P and HYAL2 in the Chinese population. Front Genet. 2020;11:977. Article  CAS  PubMed  PubMed Central  Google

Scholar  * Hachiya T, Furukawa R, Shiwa Y, Ohmomo H, Ono K, Katsuoka F, et al. Genome-wide identification of inter-individually variable DNA methylation sites improves the efficacy of

epigenetic association studies. NPJ Genom Med. 2017;2:11. Article  PubMed  PubMed Central  CAS  Google Scholar  * Komaki S, Shiwa Y, Furukawa R, Hachiya T, Ohmomo H, Otomo R, et al. iMETHYL:

an integrative database of human DNA methylation, gene expression, and genomic variation. Hum Genome Var. 2018;5:18008. Article  CAS  PubMed  PubMed Central  Google Scholar  * Richardson B.

Impact of aging on DNA methylation. Ageing Res. Rev. 2003;2:245–61. Article  CAS  PubMed  Google Scholar  * Zhang X, Ho SM. Epigenetics meets endocrinology. J Mol Endocrinol.

2011;46:R11–32. Article  CAS  PubMed  PubMed Central  Google Scholar  * Dunneram Y, Greenwood DC, Cade JE. Diet, menopause and the risk of ovarian, endometrial and breast cancer. Proc Nutr

Soc. 2019;78:438–48. Article  PubMed  Google Scholar  * Houseman EA, Accomando WP, Koestler DC, Christensen BC, Marsit CJ, Nelson HH, et al. DNA methylation arrays as surrogate measures of

cell mixture distribution. BMC Bioinforma. 2012;13:86. Article  Google Scholar  * Talens RP, Boomsma DI, Tobi EW, Kremer D, Jukema JW, Willemsen G, et al. Variation, patterns, and temporal

stability of DNA methylation: considerations for epigenetic epidemiology. FASEB J. 2010;24:3135–44. Article  CAS  PubMed  Google Scholar  * Engel AG. Genetic basis and phenotypic features of

congenital myasthenic syndromes. Handb Clin Neurol. 2018;148:565–89. Article  PubMed  Google Scholar  * Ravenscroft G, Sollis E, Charles AK, North KN, Baynam G, Laing NG. Fetal akinesia:

review of the genetics of the neuromuscular causes. J Med Genet. 2011;48:793–1. Article  CAS  PubMed  Google Scholar  * Zhuang QS, Zheng H, Gu XD, Shen L, Ji HF. Detecting the genetic link

between Alzheimer’s disease and obesity using bioinformatics analysis of GWAS data. Oncotarget. 2017;8:55915–9. Article  PubMed  PubMed Central  Google Scholar  * Aittaleb M, Chen PJ,

Akaaboune M. Failure of lysosome clustering and positioning in the juxtanuclear region in cells deficient in rapsyn. J Cell Sci. 2015;128:3744–56. CAS  PubMed  PubMed Central  Google Scholar

  * Gautam M, Noakes PG, Mudd J, Nichol M, Chu GC, Sanes JR, et al. Failure of postsynaptic specialization to develop at neuromuscular junctions of rapsyn-deficient mice. Nature.

1995;377:232–6. Article  CAS  PubMed  Google Scholar  * Lee CH, Huang CS, Chen CS, Tu SH, Wang YJ, Chang YJ, et al. Overexpression and activation of the alpha9-nicotinic receptor during

tumorigenesis in human breast epithelial cells. J Natl Cancer Inst. 2010;102:1322–35. Article  CAS  PubMed  Google Scholar  * Key TJ, Chen J, Wang DY, Pike MC, Boreham J. Sex hormones in

women in rural China and in Britain. Br J Cancer. 1990;62:631–6. Article  CAS  PubMed  PubMed Central  Google Scholar  * Parida S, Sharma D. The Microbiome-Estrogen Connection and Breast

Cancer Risk. Cells. 2019;8. https://doi.org/10.3390/cells8121642. * Prall OW, Rogan EM, Sutherland RL. Estrogen regulation of cell cycle progression in breast cancer cells. J Steroid Biochem

Mol Biol. 1998;65:169–74. Article  CAS  PubMed  Google Scholar  * Thomas T, Thomas TJ. Regulation of cyclin B1 by estradiol and polyamines in MCF-7 breast cancer cells. Cancer Res.

1994;54:1077–84. CAS  PubMed  Google Scholar  * Xing G, Xiong WC, Mei L. Rapsyn as a signaling and scaffolding molecule in neuromuscular junction formation and maintenance. Neurosci Lett.

2020;731:135013. Article  CAS  PubMed  Google Scholar  * Brack AS, Conboy MJ, Roy S, Lee M, Kuo CJ, Keller C, et al. Increased Wnt signaling during aging alters muscle stem cell fate and

increases fibrosis. Science. 2007;317:807–10. Article  CAS  PubMed  Google Scholar  * Ono M, Yin P, Navarro A, Moravek MB, Coon JS, Druschitz SA, et al. Paracrine activation of

WNT/beta-catenin pathway in uterine leiomyoma stem cells promotes tumor growth. Proc Natl Acad Sci USA. 2013;110:17053–8. Article  CAS  PubMed  PubMed Central  Google Scholar  * Seto JT,

Chan S, Turner N, MacArthur DG, Raftery JM, Berman YD, et al. The effect of alpha-actinin-3 deficiency on muscle aging. Exp Gerontol. 2011;46:292–302. Article  CAS  PubMed  Google Scholar  *

Menard S, Tagliabue E, Campiglio M, Pupa SM. Role of HER2 gene overexpression in breast carcinoma. J Cell Physiol. 2000;182:150–62. Article  CAS  PubMed  Google Scholar  * Slamon DJ, Clark

GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science. 1987;235:177–82. Article  CAS 

PubMed  Google Scholar  * Simonova OA, Kuznetsova EB, Tanas AS, Rudenko VV, Poddubskaya EV, Kekeeva TV, et al. Abnormal Hypermethylation of CpG Dinucleotides in Promoter Regions of Matrix

Metalloproteinases Genes in Breast Cancer and Its Relation to Epigenomic Subtypes and HER2 Overexpression. Biomedicines. 2020;8. https://doi.org/10.3390/biomedicines8050116. * Fiegl H,

Millinger S, Goebel G, Muller-Holzner E, Marth C, Laird PW, et al. Breast cancer DNA methylation profiles in cancer cells and tumor stroma: association with HER-2/neu status in primary

breast cancer. Cancer Res. 2006;66:29–33. Article  CAS  PubMed  Google Scholar  * Park HL. Epigenetic Biomarkers for Environmental Exposures and Personalized Breast Cancer Prevention. Int J

Environ Res Public Health. 2020;17. https://doi.org/10.3390/ijerph17041181. * Kadayifci FZ, Zheng S, Pan YX. Molecular Mechanisms Underlying the Link between Diet and DNA Methylation. Int J

Mol Sci. 2018;19. https://doi.org/10.3390/ijms19124055. * Chen N, Wang J. Wnt/beta-catenin signaling and obesity. Front Physiol. 2018;9:792. Article  PubMed  PubMed Central  Google Scholar 

* Debebe A, Medina V, Chen CY, Mahajan IM, Jia C, Fu D, et al. Wnt/beta-catenin activation and macrophage induction during liver cancer development following steatosis. Oncogene.

2017;36:6020–9. Article  CAS  PubMed  PubMed Central  Google Scholar  Download references ACKNOWLEDGEMENTS This work was funded by the National Natural Science Foundation of China (Grant

Nos. 81803303 and 81874122), the Natural Science Foundation of Jiangsu Province (Grant No. BK20180674), the CAMS Initiative for Innovative Medicine (Grant No. 2017-I2M-3-004), the Key

Program of Science and Technology Department of Sichuan Province (Grant No. 2019YFS0377), and the Jiangsu Funding (WSW-009). AUTHOR INFORMATION Author notes * These authors contributed

equally: Shuifang Lei, Lixi Li AUTHORS AND AFFILIATIONS * Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing, China Shuifang Lei, 

Qiming Yin & Rongxi Yang * Department of Medical Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and

Peking Union Medical College, Beijing, China Lixi Li & Fei Ma * Department of Breast Surgery, West China Hospital, Sichuan University, Chengdu, China Xiaoqin Yang * Department of

Clinical Laboratory, Jiangsu Province Hospital of Chinese Medicine, Nanjing, China Tian Xu & Wanjian Gu * Chengdu Shang Jin Nan Fu Hospital, West China Hospital, Sichuan University,

Chengdu, China Wenjie Zhou Authors * Shuifang Lei View author publications You can also search for this author inPubMed Google Scholar * Lixi Li View author publications You can also search

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search for this author inPubMed Google Scholar * Tian Xu View author publications You can also search for this author inPubMed Google Scholar * Wenjie Zhou View author publications You can

also search for this author inPubMed Google Scholar * Wanjian Gu View author publications You can also search for this author inPubMed Google Scholar * Fei Ma View author publications You

can also search for this author inPubMed Google Scholar * Rongxi Yang View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS The first draft of

the manuscript was written by RY and SL. RY and FM supervised and design the project. SL and QY performed the experiments and analyzed the results. LL, XY, TX, WZ, WG, and FM provided the

materials and supervised the patient enrollment and acquisition of biological samples and clinical data. All authors read and approved the final manuscript. CORRESPONDING AUTHORS

Correspondence to Fei Ma or Rongxi Yang. ETHICS DECLARATIONS CONFLICT OF INTEREST The authors declare no competing interests. ETHICAL APPROVAL All procedures were in accordance with the

ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The studies

involving human participants were reviewed and approved by the Ethics Committee of Nanjing Medical University, the Ethics Committee of Chinese Academy of Medical Sciences, and the Ethics

Committee of West China Hospital. INFORMED CONSENT The patients/participants provided their written informed consent to participate in this study. ADDITIONAL INFORMATION PUBLISHER’S NOTE

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RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Lei, S., Li, L., Yang, X. _et al._ The association between _RAPSN_ methylation in peripheral blood and

breast cancer in the Chinese population. _J Hum Genet_ 66, 1069–1078 (2021). https://doi.org/10.1038/s10038-021-00933-x Download citation * Received: 02 February 2021 * Revised: 15 April

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