Department of Obstetrics and Gynecology, Donghu District, the Second Affiliated Hospital of Nanchang University, No. 1, Minde Road, Nanchang, 330006, Jiangxi Province, China.
Mol Cell Biochem. 2020 Dec;475(1-2):185-194. doi: 10.1007/s11010-020-03871-y. Epub 2020 Aug 18.
Endometriosis is a common, chronic and painful disease in women, whose pathogenesis remains not entirely clear. Long non-coding RNA (lncRNA) MALAT1 participates in the development of endometriosis. This study further investigated the regulation of MALAT1-miR-126-5p-CREB1 axis in the pathological process of endometriosis. MALAT1, miR-126-5p, and CREB1 levels in human endometrial stromal cells (HESCs) were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Protein levels were determined by Western blotting. Cell viability and apoptosis was assessed by MTT assay and annexin V-FITC staining, respectively. The interactivity between miR-126-5p and MALAT1 (or CREB1) was assessed by dual luciferase reporter system. Knockdown of MALAT1 or CREB1 restrained proliferation and induced apoptosis as confirmed by upregulating cleaved caspase-3 and Bax, and down-regulating Bcl-2 in HESCs, while inhibition of miR-126-5p presented the opposite results. Moreover, silencing of MALAT1 triggered apoptosis of HESCs via targeting miR-126-5p. In addition, miR-126-5p directly regulated CREB1 expression via binding to its 3' non-coding region. Finally, miR-126-5p inhibitor-mediated apoptosis inhibition was restrained by CREB1 silencing via inactivation of PI3K-AKT pathway in HESCs. Taken together, our study firstly demonstrates that MALAT1 regulates apoptosis of HESCs through miR-126-5p/CREB1 axis mediated PI3K/AKT pathway. Our findings explained the pathogenesis of endometriosis and offered promising therapeutic option for endometriosis.
子宫内膜异位症是一种常见的、慢性的、疼痛的妇女疾病，其发病机制尚不完全清楚。长链非编码 RNA（lncRNA）MALAT1 参与子宫内膜异位症的发展。本研究进一步探讨了 MALAT1-miR-126-5p-CREB1 轴在子宫内膜异位症病理过程中的调节作用。通过实时定量逆转录聚合酶链反应（RT-qPCR）检测人子宫内膜基质细胞（HESCs）中 MALAT1、miR-126-5p 和 CREB1 的水平。通过 Western blot 测定蛋白水平。通过 MTT 测定和 Annexin V-FITC 染色分别评估细胞活力和细胞凋亡。通过双荧光素酶报告系统评估 miR-126-5p 与 MALAT1（或 CREB1）的相互作用。沉默 MALAT1 可通过上调 cleaved caspase-3 和 Bax，下调 HESCs 中的 Bcl-2，从而抑制细胞增殖并诱导细胞凋亡，而抑制 miR-126-5p 则呈现相反的结果。此外，沉默 MALAT1 通过靶向 miR-126-5p 触发 HESCs 凋亡。此外，miR-126-5p 通过结合其 3'非编码区直接调节 CREB1 的表达。最后，通过 PI3K-AKT 通路失活，miR-126-5p 抑制剂介导的细胞凋亡抑制被 CREB1 沉默所抑制。总之，本研究首次证明 MALAT1 通过 miR-126-5p/CREB1 轴调节 PI3K/AKT 通路调节 HESCs 凋亡。我们的研究结果解释了子宫内膜异位症的发病机制，并为子宫内膜异位症提供了有前途的治疗选择。
