Increased N6-methyladenosine causes infertility is associated with FTO expression
Increased N6-methyladenosine causes infertility is associated with FTO expression
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DOI：
10.1002/jcp.26507
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发表时间：
2018-09-01
影响因子：
5.6
通讯作者：
Huang, Boxian
中科院分区：
文献类型：
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作者：
Ding, Chenyue;Zou, Qinyan;Huang, Boxian
N6-甲基腺苷(M6A)修饰在哺乳动物转录组的表观遗传调控中起着核心作用。M6A可以被脂肪质量和肥胖相关(FTO)蛋白和酮戊二酸依赖的双加氧酶碱性B同源5(ALKBH5)蛋白去甲基化。至于m6A含量是否与卵巢功能不全(POI)疾病有关，人们知之甚少。在这项病例对照研究中，69例POI患者和53例输卵管闭塞患者来自本院生殖中心。在POI动物模型实验中，取10只POI小鼠和9只健康小鼠的卵巢组织。建立m6A检测试剂盒，用定量聚合酶链式反应和蛋白质印迹法检测FTO和ALKBH5的mRNA和蛋白表达水平。用流式细胞仪检测细胞增殖和凋亡水平，用siRNA建立FTO和ALKBH5基因敲除细胞系。结果表明，POI患者和POI小鼠的RNA中m6A含量均显著高于对照组，POI以m6A含量为特征。POI组FTO的mRNA和蛋白表达水平明显低于对照组，且与POI的危险性相关。提示FTO基因表达水平的降低可能与POI中m6A的表达增加有关，从而进一步增加POI并发症的风险。高m6A作为POI的一个新的潜在生物标志物值得进一步研究。
The N6-methyladenosine (m6A) modification plays a central role in epigenetic regulation of the mammalian transcriptome. m6A can be demethylated by the fat mass- and obesity-associated (FTO) protein and the -ketoglutarate-dependent dioxygenase alkB homolog 5 (ALKBH5) protein. Much less is known about that whether m6A content is involved in POI (premature ovarian insufficiency) disease. In this case-controlled study, 69 POI and 53 tubal occlusion patients were recruited from the reproduction centers in our hospital. For the POI animal model experiment, ovarian tissue was obtained from ten POI and nine healthy mice. An m6A test kit was developed to determine the m6A content in the RNA, and qPCR and western blot were used to examine the mRNA and protein expression levels of FTO and ALKBH5. FACS was used to measure the levels of proliferation and apoptosis, and siRNA was used to establish FTO and ALKBH5 knockdown cell lines. Our results showed that the m6A content in the RNA from POI patients and POI mice was significantly higher than control groups and that POI was characterized by the content of m6A. The mRNA and protein expression levels of FTO were significantly lower in the POI patients than control group and were associated with a risk of POI. These data suggest that the decreased mRNA and protein expression levels of FTO may be responsible for the increase in m6A in POI, which may further increase the risk of complications of POI. High m6A should be investigated further as a novel potential biomarker of POI.