APEX1在各型子宫内膜异位症中的表达及在腹膜型子宫内膜异位发生及发展中的功能研究

The exact pathogenesis of endometriosis is complecated. APEX1 is a multi-function protein involved in both DNA repair and oxidative stress response related pathways. Our previous study indicated that the expression of APEX1 is higher endometriosis tissue samples, and extremely high in the samples from patients with peritoneal endometriosis. We also examined the expression of 9 candidate miRNAs and find significant positive correlations between miR-24, miR-196b and APEX1. We intend to study the function of APEX1 from the following three aspects. (1) Detect the expression of APEX1 in different types of endometriosis and establish the relationship between APEX1 and endometriosis subtypes. (2) Examine differentially expressed miRNAs in ectopic tissues and analyze the correlation between APEX1 and miRNAs, and subsequently establish the relationship between the expression of APEX1 and miRNAs in endometriotic tissues. (3) Identify the target genes of candidate miRNAs and investigate their biological functions in vitro. Partially unveil the relationship between APEX1 and the occurrence, development and cancerization of endometriosis. Depending on the above studies, we will reveal the role of APEX1 in the occurrence and development of endometriosis, providing a tool for clinical classification and individualized medical treatment.

子宫内膜异位症致病因素复杂。APEX1是一个多功能蛋白，既参与DNA修复，也在氧化应激过程中发挥着重要的作用。我们前期研究发现APEX1的表达量在异位内膜组织中表达上升，且以腹膜型异位组织为最高。我们同时检测了9个候选miRNAs在异位子宫内膜中的表达，发现miR-24和miR-196b与APEX1的表达存在显著的相关性。我们拟研究：（1）检测APEX1在不同类型子宫内膜异位组织中的表达量，建立APEX1与子宫内膜异位分型之间的关系。（2）检测异位组织中差异表达的miRNAs，分析APEX1与miRNAs表达的相关性，建立APEX1与miRNAs表达量在子宫内膜异位组织中的关系。（3）鉴定差异miRNAs的靶基因，并在细胞水平验证其生物学功能，阐述APEX1与子宫内膜异位发生、发展及癌变的关系。以揭示APEX1在子宫内膜异位发生、发展中的作用。

子宫内膜异位症（内异症）是严重影响育龄期妇女健康的疾病。APEX1在细胞分化、增殖和凋亡中起转录共激活剂作用。目前的研究仅限于将 APEX1 与子宫内膜异位症发展联系起来的假定机制，但对 APEX1 在子宫内膜异位症病变中的潜在调节功能和下游靶点尚无研究。miR-24 的表达水平与子宫内膜异位症有一定的关系。本研究主要研究内容及结果：.1.APEX1的表达量在子宫内膜异位组织中显著高于正常组织，且以腹膜型异位组织为最高。 .2.原代子宫内膜间质细胞vimentin的阳性率96％，纯度和活力良好，可用于下一步实验。.3.APEX1加速子宫内膜间质细胞增殖并抑制细胞凋亡。.4.miR-24在PE组织中强烈表达，APEX1与miR-24的表达呈正相关。.5.miR-24对子宫内膜间质细胞的调控作用：与mimic NC组相比，转染miR-24 mimic，子宫内膜间质细胞染色阳性细胞数明显增多显示细胞增殖活力显著升高；与inhibitor-NC组相比，转染miR-24 inhibitor后，子宫内膜间质细胞染色阳性细胞数明显减少，显示细胞增殖活力显著下降。转染miR-24 mimic，子宫内膜间质细胞凋亡水平显著下降，与inhibitor NC组相比，转染miR-24 inhibitor后，异位子宫内膜间质细胞凋亡水平水平显著升高。与mimic-NC组相比，miR-24 mimic组BAX、cleaved-casase-3的表达水平显著下降，BCL-2的表达水平显著上升；与inhibitor-NC相比，miR-24 inhibitor组BAX、cleaved-casase-3的表达水平显著上升，BCL-2的表达水平显著下降。.6.APEX1与miR-24之间的调控作用：证明APEX1是miR-24的上游调控分子，RIP检测APEX1和pri-miR-24的结合：与转染空载体组相比，检测到过表达APEX1组中获得的pri-miR-24显著增加，说明APEX1可以直接与pri-miR-24结合。在细胞中转染sh-APEX1，检测到pri-miR-24水平增加，结果提示，敲低APEX1的表达水平，导致pri-miR-24合成miR-24这个过程受到损害，从而pri-miR-24产生蓄积。.这些发现表明 APEX1 可以上调 miR-24 以作为子宫内膜异位症的新治疗靶点。