Altered expression of microRNA-451 in eutopic endometrium of baboons (Papio anubis) with endometriosis
Altered expression of microRNA-451 in eutopic endometrium of baboons (Papio anubis) with endometriosis
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DOI：
10.1093/humrep/dev229
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发表时间：
2015-12-01
影响因子：
6.1
通讯作者：
Fazleabas, A. T.
中科院分区：
文献类型：
--
作者：
Joshi, N. R.;Su, R. W.;Fazleabas, A. T.
研究问题：诱导子宫内膜异位症后，狒狒异位子宫内膜（EuE）中的microrna （miRs）是否发生改变？结论：子宫内膜异位症的诱发早在疾病诱发后3个月，就会导致狒狒子宫内膜中包括miR-451在内的8种mir的表达发生显著变化。已知情况：子宫内膜异位症是最常见的妇科疾病之一，可导致育龄妇女慢性盆腔疼痛和不孕症。miRs在女性中的表达改变已被报道，并被认为在包括子宫内膜异位症在内的几种妇科疾病的病理生理中发挥重要作用。研究设计、大小、持续时间：在子宫内膜异位症诱导前和诱导后3个月，从同一组狒狒中获得EuE。在疾病诱导后3至15个月的其他狒狒的异位和异位子宫内膜中验证了miR-451表达的改变。对有或无子宫内膜异位症的女性进行定时子宫内膜活检，以验证miR-451的表达。参与者/材料，设置，方法：从诱导子宫内膜异位症前后的EuE样品中提取总RNA，并使用a8 × 15 K miR芯片分析miRNA的表达。采用AgiMiRna软件对微阵列信号数据进行预处理，并采用经验贝叶斯模型估计变化。本研究的重点是对微阵列数据进行定量RT-PCR验证，特别是在狒狒（n = 3）和女性[对照组（n = 7）和子宫内膜异位症（n = 19）]中的miR-451及其靶基因。使用原位杂交和免疫组织化学对miR-451和靶基因表达进行描述性和相关性分析，而使用体外3'非翻译区（UTR）荧光素酶测定和miR-451在人子宫内膜和子宫内膜异位症细胞系中的过表达进行功能分析。主要结果和CHANCE的作用：子宫内膜异位症的诱导导致狒狒子宫内膜miR-451、-141、-29c、-21、-424、-19b、-200a和-181a的表达改变。在狒狒中，子宫内膜异位症的诱导在3个月时显著降低了miR-451的表达（P < 0.001），这也与其靶基因YWHAZ的表达增加有关（14.3.3 zeta）。在子宫内膜异位症患者中，miR-451表达也有类似的显著降低（P < 0.0001）。3' UTR荧光素酶检测证实了miR-451对YWHAZ表达的调控。此外，miR-451在12Z细胞（永生化的人子宫内膜异位症上皮细胞系）中的过表达导致其靶点YWHAZ的表达降低，这与细胞增殖降低相关。局限性，谨慎原因：该研究仅关注miR-451及其靶点之一，即YWHAZ。单个miR可靶向多个基因，单个基因也可受miR数量的调控；因此，其他mir及其调控基因可能参与子宫内膜异位症的病理生理。研究结果的更广泛意义：我们的数据表明，狒狒异位病变的存在可导致EuE miR表达的变化，最早可在疾病诱发后3个月发生，其中一些变化可能持续整个疾病过程。我们提出，在狒狒和子宫内膜异位症女性中，miR-451的显著下调增加了多个靶基因的表达。其中一个靶基因YWHAZ的表达增加，增加了增殖，可能有助于疾病的病理生理。
STUDY QUESTION: Are microRNAs (miRs) altered in the eutopic endometrium (EuE) of baboons following the induction of endometriosis?SUMMARY ANSWER: Induction of endometriosis causes significant changes in the expression of eight miRs, including miR-451, in the baboon endometrium as early as 3 months following induction of the disease.WHAT IS KNOWN ALREADY: Endometriosis is one of the most common gynecological disorders and causes chronic pelvic pain and infertility in women of reproductive age. Altered expression of miRs has been reported in women and has been suggested to play an important role in the pathophysiology of several gynecological disorders including endometriosis.STUDY DESIGN, SIZE, DURATION: EuE was obtained from the same group of baboons before and 3 months after the induction of endometriosis. The altered expression of miR-451 was validated in the eutopic and ectopic endometrium of additional baboons between 3 and 15 months following disease induction. Timed endometrial biopsies from women with and without endometriosis were also used to validate the expression of miR-451.PARTICIPANTS/MATERIALS, SETTING, METHODS: Total RNA was extracted from EuE samples before and after the induction of endometriosis, and miRNA expression was analyzed using a8 x 15 K miR microarray. Microarray signal data were preprocessed by AgiMiRna software, and an empirical Bayes model was used to estimate the changes. The present study focused on quantitative RT-PCR validation of the microarray data, specifically on miR-451 and its target genes in both baboons (n = 3) and women [control (n = 7) and endometriosis (n = 19)]. Descriptive and correlative analysis of miR-451 and target gene expression was conducted using in situ hybridization and immunohistochemistry, while functional analysis utilized an in vitro 3' untranslated region (UTR) luciferase assay and overexpression of miR-451 in human endometrial and endometriotic cell lines.MAIN RESULTS AND THE ROLE OF CHANCE: Induction of endometriosis results in the altered expression of miR-451, -141, -29c, -21, -424, -19b, -200a and -181a in the baboon endometrium. In the baboon, induction of endometriosis significantly decreased the expression of miR-451 at 3 months (P < 0.001), which was also associated with increased expression of its target gene YWHAZ (14.3.3 zeta). A similar significant (P < 0.0001) decrease in miR-451 expression was observed in women with endometriosis. The 3' UTR luciferase assay confirmed the regulation of YWHAZ expression by miR-451. Furthermore, overexpression of miR-451 in 12Z cells (immortalized human endometriotic epithelial cell line) led to the decreased expression of its target YWHAZ and this was correlated with decreased cell proliferation.LIMITATIONS, REASONS FOR CAUTION: The study focused only on miR-451 and one of its targets, namely YWHAZ. A single miR could target number of genes and a single gene could also be regulated by number of miRs; hence, it is possible that other miRs and their regulated genes may contribute to the pathophysiology of endometriosis.WIDER IMPLICATIONS OF THE FINDINGS: Our data suggest that the presence of ectopic lesions in baboon causes changes in EuE miR expression as early as 3 months postinduction of the disease, and some of these changes may persist throughout the course of the disease. We propose that the marked down-regulation of miR-451 in both baboons and women with endometriosis increases the expression of multiple target genes. Increased expression of one of the target genes, YWHAZ, increases proliferation, likely contributing to the pathophysiology of the disease.