High-resolution Melting Analysis of MED12 Mutations in Uterine Leiomyomas in Chinese Patients

Overview

Journal Genet Test Mol Biomarkers

Specialties Genetics
Molecular Biology

Date 2015 Jan 24

PMID 25615570

Citations 9

Authors

Hua Wang

Jun Ye

Hua Qian

Ruifang Zhou

Jun Jiang

Lihua Ye

Affiliations

Soon will be listed here.

Abstract

Objectives: Somatic mutations in mediator complex subunit 12 (MED12) have emerged as a critical genetic change in the development of uterine leiomyomas. Studies, however, have focused largely on cohorts consisting of Caucasian patients. In this study, uterine leiomyomas from Chinese patients were examined for MED12 mutations. In addition, polymerase chain reaction (PCR)-based high-resolution melting analysis (HRMA) was compared with direct sequencing as a potentially more sensitive method for the detection of MED12 mutations.

Methods: Tissue samples with the pathologies of uterine leiomyoma (n=181) and other endometrial diseases (n=157) were collected from Chinese patients at the Taizhou People's Hospital and Taizhou Polytechnic College (Taizhou City, China). Genomic DNA was prepared from all samples. Both PCR-based HRMA and PCR-based direct sequencing were used to detect MED12 mutations.

Results: PCR-based HRMA and direct sequencing revealed MED12 mutations in 95/181 (52.5%) and 93/181 (51.4%) uterine leiomyomas, respectively. Nearly half of these mutations (46/93) were found in a single codon, codon 131. The coincidence rate between the two methods was 98.9% (179/181) so that no statistically significant difference was evident in the application of the methodologies (χ(2)=0.011, p=0.916). In addition, MED12 mutations were identified in 1/157 (4.17%) case of other endometrial pathologies by both methods.

Conclusions: MED12 mutations were closely associated with the development of uterine leiomyomas, as opposed to other uterine pathologies in Chinese patients, and PCR-based HRMA was found to be a reliable method for the detection of MED12 mutations.

Citing Articles

Comprehensive Review of Uterine Fibroids: Developmental Origin, Pathogenesis, and Treatment.

Yang Q, Ciebiera M, Bariani M, Ali M, Elkafas H, Boyer T Endocr Rev. 2021; 43(4):678-719.

PMID: 34741454 PMC: 9277653. DOI: 10.1210/endrev/bnab039.

Frequency of MED12 Mutation in Relation to Tumor and Patient's Clinical Characteristics: a Meta-analysis.

He C, Nelson W, Li H, Xu Y, Dai X, Wang Y Reprod Sci. 2021; 29(2):357-365.

PMID: 33569750 DOI: 10.1007/s43032-021-00473-x.

Understanding Obesity as a Risk Factor for Uterine Tumors Using Drosophila.

Li X, Liu M, Ji J Adv Exp Med Biol. 2019; 1167:129-155.

PMID: 31520353 DOI: 10.1007/978-3-030-23629-8_8.

mutations and fumarate hydratase inactivation in uterine adenomyomas.

Heikkinen T, Ayravainen A, Hanninen J, Ahvenainen T, Butzow R, Pasanen A Hum Reprod Open. 2019; 2018(4):hoy020.

PMID: 30895261 PMC: 6276694. DOI: 10.1093/hropen/hoy020.

Mediator Kinase Disruption in MED12-Mutant Uterine Fibroids From Hispanic Women of South Texas.

Park M, Shen H, Kim N, Gao F, Failor C, Knudtson J J Clin Endocrinol Metab. 2018; 103(11):4283-4292.

PMID: 30099503 PMC: 6194812. DOI: 10.1210/jc.2018-00863.

References

Kampjarvi K, Makinen N, Kilpivaara O, Arola J, Heinonen H, Bohm J . Somatic MED12 mutations in uterine leiomyosarcoma and colorectal cancer. Br J Cancer. 2012; 107(10):1761-5. PMC: 3493861. DOI: 10.1038/bjc.2012.428. View

Huang S, Holzel M, Knijnenburg T, Schlicker A, Roepman P, McDermott U . MED12 controls the response to multiple cancer drugs through regulation of TGF-β receptor signaling. Cell. 2012; 151(5):937-50. PMC: 3672971. DOI: 10.1016/j.cell.2012.10.035. View

Matsubara A, Sekine S, Yoshida M, Yoshida A, Taniguchi H, Kushima R . Prevalence of MED12 mutations in uterine and extrauterine smooth muscle tumours. Histopathology. 2013; 62(4):657-61. DOI: 10.1111/his.12039. View

Wu S, de Borsetti N, Bain E, Bulow C, Gamse J . Mediator subunit 12 coordinates intrinsic and extrinsic control of epithalamic development. Dev Biol. 2013; 385(1):13-22. DOI: 10.1016/j.ydbio.2013.10.023. View

Turunen M, Spaeth J, Keskitalo S, Park M, Kivioja T, Clark A . Uterine leiomyoma-linked MED12 mutations disrupt mediator-associated CDK activity. Cell Rep. 2014; 7(3):654-60. PMC: 4041330. DOI: 10.1016/j.celrep.2014.03.047. View

Je E, Kim M, Min K, Yoo N, Lee S . Mutational analysis of MED12 exon 2 in uterine leiomyoma and other common tumors. Int J Cancer. 2012; 131(6):E1044-7. DOI: 10.1002/ijc.27610. View

Zhang H, Emmons S . A C. elegans mediator protein confers regulatory selectivity on lineage-specific expression of a transcription factor gene. Genes Dev. 2000; 14(17):2161-72. PMC: 316889. DOI: 10.1101/gad.814700. View

Moghal N, Sternberg P . A component of the transcriptional mediator complex inhibits RAS-dependent vulval fate specification in C. elegans. Development. 2002; 130(1):57-69. DOI: 10.1242/dev.00189. View

Chou L, Lyon E, Wittwer C . A comparison of high-resolution melting analysis with denaturing high-performance liquid chromatography for mutation scanning: cystic fibrosis transmembrane conductance regulator gene as a model. Am J Clin Pathol. 2005; 124(3):330-8. DOI: 10.1309/BF3M-LJN8-J527-MWQY. View

10.

Kim S, Xu X, Hecht A, Boyer T . Mediator is a transducer of Wnt/beta-catenin signaling. J Biol Chem. 2006; 281(20):14066-75. DOI: 10.1074/jbc.M602696200. View

11.

Philibert R, Madan A . Role of MED12 in transcription and human behavior. Pharmacogenomics. 2007; 8(8):909-16. DOI: 10.2217/14622416.8.8.909. View

12.

Ding N, Zhou H, Esteve P, Chin H, Kim S, Xu X . Mediator links epigenetic silencing of neuronal gene expression with x-linked mental retardation. Mol Cell. 2008; 31(3):347-59. PMC: 2583939. DOI: 10.1016/j.molcel.2008.05.023. View

13.

Tutter A, Kowalski M, Baltus G, Iourgenko V, Labow M, Li E . Role for Med12 in regulation of Nanog and Nanog target genes. J Biol Chem. 2008; 284(6):3709-18. DOI: 10.1074/jbc.M805677200. View

14.

Tindall E, Petersen D, Woodbridge P, Schipany K, Hayes V . Assessing high-resolution melt curve analysis for accurate detection of gene variants in complex DNA fragments. Hum Mutat. 2009; 30(6):876-83. DOI: 10.1002/humu.20919. View

15.

Whittall R, Scartezini M, Li K, Hubbart C, Reiner Z, Abraha A . Development of a high-resolution melting method for mutation detection in familial hypercholesterolaemia patients. Ann Clin Biochem. 2009; 47(Pt 1):44-55. DOI: 10.1258/acb.2009.009076. View

16.

Taatjes D . The human Mediator complex: a versatile, genome-wide regulator of transcription. Trends Biochem Sci. 2010; 35(6):315-22. PMC: 2891401. DOI: 10.1016/j.tibs.2010.02.004. View

17.

Makinen N, Mehine M, Tolvanen J, Kaasinen E, Li Y, Lehtonen H . MED12, the mediator complex subunit 12 gene, is mutated at high frequency in uterine leiomyomas. Science. 2011; 334(6053):252-5. DOI: 10.1126/science.1208930. View

18.

Makinen N, Heinonen H, Moore S, Tomlinson I, van der Spuy Z, Aaltonen L . MED12 exon 2 mutations are common in uterine leiomyomas from South African patients. Oncotarget. 2011; 2(12):966-9. PMC: 3282101. DOI: 10.18632/oncotarget.370. View

19.

McGuire M, Yatsenko A, Hoffner L, Jones M, Surti U, Rajkovic A . Whole exome sequencing in a random sample of North American women with leiomyomas identifies MED12 mutations in majority of uterine leiomyomas. PLoS One. 2012; 7(3):e33251. PMC: 3299761. DOI: 10.1371/journal.pone.0033251. View