APEX1/miR-24 轴：子宫内膜异位症治疗的新靶点。
APEX1/miR-24 axis: a promising therapeutic target in endometriosis.
机构信息
Department of Obstetrics and Gynecology, Wuhan University Renmin Hospital, Wuhan, 430060, Hubei, China.
Department of Oncology, Wuhan University Renmin Hospital, No. 99, Zhangzhidong Road, Wuchang District, Wuhan, 430060, Hubei, China.
出版信息
Arch Gynecol Obstet. 2021 Jul;304(1):131-141. doi: 10.1007/s00404-021-05963-6. Epub 2021 Jan 27.
PURPOSE
The present work aimed to explore the aberrant expression of APEX1 in endometrial stromal cells (ESC) and the underlying mechanisms.
METHODS
The levels of APEX1 and miR-24 in endometriosis tissues were tested by qRT-PCR and Western blot. After cell transfection, cells were correspondingly classified into pcDNA3.1-NC, sh-NC, mimic NC, inhibitor NC, pcDNA3.1-APEX1, sh-APEX1, miR-24 mimic, miR-24 inhibitor, sh-NC + inhibitor NC, inhibitor-NC + sh-APEX1, sh-NC + miR-24 inhibitor, pcDNA3.1-NC + mimic NC, mimic NC + pcDNA3.1-APEX1 and pcDNA3.1-NC + miR-24 mimic group. Besides, cell proliferation, apoptosis in addition to apoptosis-related proteins Bax, Bcl-2 and cleaved-casase-3 were analyzed by BrdU assay, flow cytometry (FCM) and Western blot assays, respectively. Additionally, RIP assay was conducted to determine the interaction between pri-miR-24 and miR-24.
RESULTS
APEX1 and miR-24 were highly expressed in endometriosis tissues. Overexpression of APEX1 and miR-24 potentiates ESC proliferation and inhibits apoptosis, while those effects could be reversed by APEX1 and miR-24 silencing. Meanwhile, APEX1 and miR-24 could elevate ESC apoptosis-related proteins Bax and cleaved-caspase-3 and decrease Bcl-2 expression. Importantly, APEX1 was positively correlated with miR-24 expression.
CONCLUSION
APEX1 promotes ESC proliferation and inhibits apoptosis by upregulating miR-24 expression.
目的
本研究旨在探讨 APEX1 在子宫内膜基质细胞（ESC）中的异常表达及其潜在机制。
方法
采用 qRT-PCR 和 Western blot 检测子宫内膜异位症组织中 APEX1 和 miR-24 的水平。转染细胞后，将细胞相应地分为 pcDNA3.1-NC、sh-NC、mimic NC、inhibitor NC、pcDNA3.1-APEX1、sh-APEX1、miR-24 mimic、miR-24 inhibitor、sh-NC+inhibitor NC、inhibitor-NC+sh-APEX1、sh-NC+miR-24 inhibitor、pcDNA3.1-NC+mimic NC、mimic NC+pcDNA3.1-APEX1 和 pcDNA3.1-NC+miR-24 mimic 组。此外，通过 BrdU 检测、流式细胞术（FCM）和 Western blot 分析细胞增殖、凋亡以及凋亡相关蛋白 Bax、Bcl-2 和 cleaved-casase-3。另外，通过 RIP 测定确定 pri-miR-24 与 miR-24 的相互作用。
结果
APEX1 和 miR-24 在子宫内膜异位症组织中高表达。APEX1 和 miR-24 的过表达促进 ESC 增殖并抑制凋亡，而 APEX1 和 miR-24 的沉默可逆转这些作用。同时，APEX1 和 miR-24 可上调 ESC 凋亡相关蛋白 Bax 和 cleaved-caspase-3，降低 Bcl-2 表达。重要的是，APEX1 与 miR-24 的表达呈正相关。
结论
APEX1 通过上调 miR-24 的表达促进 ESC 增殖并抑制凋亡。
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