Wang Xiaoyan, Zhang Xiruo, Liu Shan, Li Guangyu, Cui Linlin, Qin Yingying, Chen Zi-Jiang
Center for Reproductive Medicine, Shandong Provincial Hospital Affiliated to Shandong University; Shandong Provincial Key Laboratory of Reproductive Medicine.
National Research Center for Assisted Reproductive Technology and Reproductive Genetics, China.
Hum Reprod. 2016 Dec;31(12):2865-2871. doi: 10.1093/humrep/dew259. Epub 2016 Oct 18.
Are mutations and/or polymorphisms in the TP63 gene associated with human Müllerian duct anomalies (MDAs)?
The novel mutation c.*374 G > A in the TP63 gene resulted in decreased expression of TP63 by generating new binding sites with miR-1260a/miR-532-3p and revealed the potential association between TP63 and human MDAs.
It has been shown that mice lacking Tp63 exhibit hypoplastic genitalia, a single cloacal opening, and persistence of columnar epithelium at lower genital tract sites. It has also been reported that a nonsense mutation in EMX2 results in decreased TP63 expression in a woman with MDAs. However, generally in humans the association between TP63 and MDAs is unknown.
STUDY DESIGN, SIZE, DURATION: A total of 200 unrelated Chinese women with MDAs and 200 unrelated Chinese women with a normal uterus and vagina, as controls, were recruited in the Center for Reproductive Medicine of Shandong University. All participants had a normal karyotype (46, XX).
PARTICIPANTS/MATERIALS, SETTING, METHODS: The 20 exons of the TP63 gene were sequenced in 200 cases and 200 controls. Putative binding sites for microRNAs were validated by dual luciferase activity assays. The role of microRNAs was further examined by western blot.
Sequence analysis revealed 15 known single-nucleotide polymorphisms. Additionally, three novel heterozygous variants, c.387 G > C, c.*374 G > A and c.*749 G > A, were identified in three patients with MDAs, none of which were detected in controls. Variant c.*374 G > A, located in the 3' untranslated region, was highly conserved among mammals and predicted to create microRNAs binding sites, which was confirmed by dual luciferase activity assays. Western blot demonstrated that the binding with miR-1260a/miR-532-3p resulting from the variation c.*374 G > A decreased the expression of TP63.
N/A LIMITATIONS, REASONS FOR CAUTION: Further study is needed to uncover the role of the EMX2-TP63 pathway in the development of the Müllerian duct.
This study revealed the possible association between TP63 and MDAs and suggested a potential contribution of microRNA-regulated expression of genes in the etiology of MDAs.
STUDY FUNDING/COMPETING INTERESTS: This research was supported by National Basic Research Program of China (973 Program) (2012CB944700), the State Key Program of National Natural Science Foundation of China (81430029), the National Natural Science Foundation of China (81270662, 81471509), China Postdoctoral Science Foundation (2014M561939) and the Scientific Research Foundation of Shandong Province of Outstanding Young Scientists (BS2014YY013, 2014BSE27022). The authors have no competing interests.
TP63基因中的突变和/或多态性与人类苗勒管异常（MDA）有关吗？
TP63基因中的新突变c.*374 G>A通过与miR-1260a/miR-532-3p产生新的结合位点导致TP63表达降低，并揭示了TP63与人类MDA之间的潜在关联。
已表明缺乏Tp63的小鼠表现出生殖器发育不全、单一泄殖腔开口以及下生殖道部位柱状上皮的持续存在。也有报道称，一名患有MDA的女性中EMX2的无义突变导致TP63表达降低。然而，在人类中，TP63与MDA之间的关联通常尚不清楚。
研究设计、规模、持续时间：山东大学生殖医学中心招募了200名患有MDA的无血缘关系的中国女性和200名子宫和阴道正常的无血缘关系的中国女性作为对照。所有参与者的核型均正常（46,XX）。
参与者/材料、设置、方法：对200例病例和200名对照的TP63基因的20个外显子进行测序。通过双荧光素酶活性测定验证了微小RNA的假定结合位点。通过蛋白质印迹进一步研究了微小RNA的作用。
序列分析揭示了15个已知的单核苷酸多态性。此外，在三名患有MDA的患者中鉴定出三个新的杂合变体，c.387 G>C、c.*374 G>A和c.*749 G>A，在对照中均未检测到。位于3'非翻译区的变体c.*374 G>A在哺乳动物中高度保守，并预测会产生微小RNA结合位点，这通过双荧光素酶活性测定得到证实。蛋白质印迹表明，由变体c.*374 G>A导致的与miR-1260a/miR-532-3p的结合降低了TP63的表达。
无
局限性、谨慎的理由：需要进一步研究以揭示EMX2-TP63通路在苗勒管发育中的作用。
本研究揭示了TP63与MDA之间的可能关联，并表明微小RNA调节的基因表达在MDA病因学中的潜在作用。
研究资金/竞争利益：本研究得到了中国国家基础研究计划（973计划）（2012CB944700）、中国国家自然科学基金重点项目（81430029）、中国国家自然科学基金（81270662,81471509）、中国博士后科学基金（2014M561939）以及山东省杰出青年科学家科研基金（BS2014YY013,2014BSE27022）的支持。作者没有竞争利益。
