CircRNA_000809通过miR-200c-3p对子宫内膜异位症间质细胞的增殖、迁移及上皮间质转化的影响

目的：探讨circRNA_000809通过靶向miR-200c-3p调控子宫内膜异位症间质细胞增殖、迁移及上皮-间质转化（EMT）的作用。方法：收集2020年1月至2022年1月于温州医科大学附属第二医院育英儿童医院行卵巢囊肿剥除术患者的卵巢巧克力囊肿组织25例，设为子宫内膜异位症组；收集25例正常子宫内膜组织，标本来自于因早期宫颈癌行子宫切除术或者行宫腔镜检查术的育龄期非子宫内膜异位症患者，设为正常子宫内膜组。采用qRT-PCR法检测正常子宫内膜、异位内膜组织中circRNA_000809、miR-200c-3p的表达量；分别采用EdU实验和Transwell实验检测异位子宫内膜间质细胞的增殖及迁移；采用蛋白质印迹法（Western blot）检测相关蛋白表达水平。结果：与正常子宫内膜组织比，异位子宫内膜中circRNA_000809表达上调（P＜0.05），而miR-200c-3p的表达下调（P＜0.05）；且Pearson相关性分析显示在异位子宫内膜组织中circRNA_000809与miR-200c-3p的表达量呈负相关（P＜0.05）；分别用siNC+inhibitor NC、si-circRNA_000809+inhibitor NC和si-circRNA_000809+miR-200c-3p inhibitor共转染异位子宫内膜间质细胞，结果显示与siNC+inhibitor NC组比，si-circRNA_000809+inhibitor NC组异位子宫内膜间质细胞增殖和迁移能力减弱，ZEB1/ZEB2的蛋白水平也降低（P＜0.05）；而si-circRNA_000809+miR-200c-3p inhibitor组异位子宫内膜间质细胞增殖、迁移能力及ZEB1/ZEB2蛋白的表达较si-circRNA_000809+inhibitor NC组均有上升，但较siNC+inhibitor NC组下降（P＜0.05）。结论：抑制circRNA_000809表达可通过上调miR-200c-3p的表达从而抑制子宫内膜异位症间质细胞的迁移及EMT。

The mechanism of circRNA_000809 in regulating endometriosis stromal cell proliferation,migration and epithelial-mesenchymal transition by targeting miR-200c-3p

Objective: To explore the role of circRNA_000809 in regulating proliferation, migration and epithelial-mesenchymal transition (EMT) of endometriosis stromal cells by targeting miR-200-3p. Methods: From January 2020 to January 2022, twenty-five cases of chocolate ovarian cyst tissues from patients undergoing ovarian cyst excision in the Second Affiliated Hospital & Yuying Children’s Hospital of Wenzhou Medical University were selected as the endometriosis group. Twenty-five normal endometrial tissue samples were collected from non-endometriosis patients of childbearing age who underwent hysterectomy or hysteroscopy for early cervical cancer and were assigned to the normal endometrial group. qRT-PCR analysis was used to detect the expression of circRNA_000809 and miR-200c-3p in ectopic endometrium tissues of endometriosis patients and normal endometrium. The 5-ethynyl-2’-deoxyuridine (EdU) assays were used to detect cell proliferation ability. Transwell migration assays were employed to monitor migration capabilities. Western blot was performed to detect protein expression. Results: The circRNA_000809 expression in ectopic endometrium tissues was increased (P<0.05), and the miR-200c-3p expression was decreased compared with normal endometrium tissues (P<0.05). Pearson’s test showed that the expression of circRNA_000809 and miR-200c-3p was negatively correlated in ectopic endometrium tissues. We co-transfected ectopic endometrial stromal cells with siNC+inhibitor NC, si-circRNA_000809+inhibitor NC and si-circRNA_000809+miR-200c-3p inhibitor, respectively. The results showed that the proliferation and migration of ectopic endometrial stromal cells were attenuated and the protein levels of ZEB1/ZEB2 were also decreased in the si-circRNA_000809+inhibitor NC group compared with the siNC+inhibitor NC group; while the proliferation and migration of ectopic endometrial stromal cells and the expression of ZEB1/ZEB2 protein were reversed to some extent in the si-circRNA_000809+miR-200c-3p inhibitor group (P<0.05). Conclusion: Silencing of circRNA_000809 inhibited EESCs proliferation, migration, and EMT through miR-200c-3p/EMT signaling.