Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, 250012, People's Republic of China.
Department of Obstetrics, Shouguang People's Hospital, Shouguang, 262700, People's Republic of China.
Int J Nanomedicine. 2022 Mar 16;17:1155-1170. doi: 10.2147/IJN.S354314. eCollection 2022.
Long non-coding RNA (lncRNA) and exosomes are implicated in endometriosis development. We measured the expression of an exosomal lncRNA, homeobox transcript antisense RNA (HOTAIR), and explored its molecular mechanism in endometriosis progression.
Expression of HOTAIR and microRNA (miR)-761 in different endometrial tissues was measured. Exosomes were isolated from a culture medium of endometrial stromal cells (ESCs). RT-qPCR was used to measure HOTAIR expression in different exosome types. CCK-8, Edu, wound healing, transwell assays, flow cytometry and tube formation were used to detect the role of exosomal HOTAIR on ESCs and human umbilical vein endothelial cells (HUVECs). The relationship among miR-761, HOTAIR, and histone deacetylase 1 (HDAC1) was verified by dual-luciferase reporter assay. ESCs were transfected with miR-761 mimics or HDAC1 small interfering RNA (si-RNA) to ascertain if alterations in expression of miR-761 or HDAC1 could reverse the effect of exosomal HOTAIR. Then, we detected the effect of the HOTAIR/miR-761/HDAC1 axis on signal transducer and activator of transcription 3 (STAT3)-mediated inflammation. In vivo experiments were conducted to verify in vitro results.
HOTAIR expression was upregulated and miR-761 expression was downregulated in ectopic endometrium tissues. HOTAIR was packaged into exosomes and transported from ESCs to surrounding cells. Exosomal HOTAIR promoted the proliferation, migration, and invasion, and inhibited the apoptosis of ESCs. Angiogenesis of HUVECs was enhanced after cultured with exosomal HOTAIR. HOTAIR acted as a competing endogenous RNA to downregulate miR-761 and increase HDAC1 expression. miR-761 overexpression or HDAC1 knockdown reversed the role of exosomal HOTAIR on ESCs and HUVECs. The HOTAIR/miR-761/HDAC1 axis could activate STAT3-related proinflammatory cytokines and stattic (inhibitor of phosphorylated-STAT3) could reverse the effect of HOTAIR on ESCs and HUVECs. In vivo experiments suggested that exosomal HOTAIR promoted the growth of endometrial lesions in vivo.
Exosomal HOTAIR promoted the progression and angiogenesis of endometriosis by regulating the miR-761/HDAC1 axis and activating STAT3-mediated inflammation in vitro and in vivo, which may provide promising treatment for endometriosis.
长链非编码 RNA（lncRNA）和外泌体与子宫内膜异位症的发展有关。我们测量了外泌体 lncRNA 同源盒转录物反义 RNA（HOTAIR）的表达，并探讨了其在子宫内膜异位症进展中的分子机制。
测量不同子宫内膜组织中 HOTAIR 和 microRNA（miR）-761 的表达。从子宫内膜基质细胞（ESCs）的培养基中分离出外泌体。RT-qPCR 用于测量不同外泌体类型中 HOTAIR 的表达。CCK-8、Edu、划痕愈合、Transwell 测定、流式细胞术和管形成用于检测外泌体 HOTAIR 对 ESCs 和人脐静脉内皮细胞（HUVECs）的作用。通过双荧光素酶报告基因测定验证 miR-761、HOTAIR 和组蛋白去乙酰化酶 1（HDAC1）之间的关系。用 miR-761 模拟物或 HDAC1 小干扰 RNA（si-RNA）转染 ESCs，以确定 miR-761 或 HDAC1 的表达变化是否可以逆转外泌体 HOTAIR 的作用。然后，我们检测了 HOTAIR/miR-761/HDAC1 轴对信号转导和转录激活因子 3（STAT3）介导的炎症的影响。进行体内实验以验证体外结果。
异位子宫内膜组织中 HOTAIR 表达上调，miR-761 表达下调。HOTAIR 被包装到外泌体中，并从 ESCs 转运到周围细胞。外泌体 HOTAIR 促进 ESCs 的增殖、迁移和侵袭，并抑制其凋亡。与外泌体 HOTAIR 共培养后，HUVECs 的血管生成增强。HOTAIR 作为竞争性内源性 RNA 下调 miR-761 并增加 HDAC1 表达。miR-761 过表达或 HDAC1 敲低逆转了外泌体 HOTAIR 对 ESCs 和 HUVECs 的作用。HOTAIR/miR-761/HDAC1 轴可以激活 STAT3 相关的促炎细胞因子，而 stattic（磷酸化-STAT3 的抑制剂）可以逆转 HOTAIR 对 ESCs 和 HUVECs 的作用。体内实验表明，外泌体 HOTAIR 在体内促进了子宫内膜病变的生长。
外泌体 HOTAIR 通过调节 miR-761/HDAC1 轴和激活 STAT3 介导的炎症在体内和体外促进子宫内膜异位症的进展和血管生成，这可能为子宫内膜异位症提供有希望的治疗方法。
