Hsa_circ_0001550 通过调节子宫内膜基质细胞的增殖和凋亡来损害蜕膜化。

Hsa_circ_0001550 impairs decidualization by regulating the proliferation and apoptosis of endometrial stromal cells.

机构信息

The First Clinical Medical College of Lanzhou University, Lanzhou Gansu, China.

The Reproductive Center, The First Hospital of Lanzhou University, Lanzhou Gansu, China; Gansu Key Laboratory of Reproductive Medicine and Embryos, Lanzhou Gansu, China.

出版信息

Reprod Biomed Online. 2023 Feb;46(2):225-233. doi: 10.1016/j.rbmo.2022.10.003. Epub 2022 Oct 11.

DOI:10.1016/j.rbmo.2022.10.003

PMID:36396534

Abstract

RESEARCH QUESTION

What is the molecular function of hsa_circ_0001550 in decidualization?

DESIGN

Human endometrial stromal cells (HESC) were isolated from the endometrium tissues to build an in-vitro decidualization model. Different concentrations of medroxyprogesterone acetate (MPA) were used to observe whether the expression level of hsa_circ_0001550 was related to progesterone. Biological characteristics and distribution of hsa_circ_0001550 were determined by RNase R, actinomycin D (Act D) assay and cytoplasmic/nuclear fraction assay. Then the overexpression of hsa_circ_0001550 was achieved by adenovirus vector. Cell proliferation was determined by Cell Counting Kit-8 (CCK-8) assays. The cell cycle was assessed by flow cytometry analyses. Cell apoptosis was determined by annexin-V/propidium iodide double staining experiment and western blotting.

RESULTS

The expression of hsa_circ_0001550 was decreased in decidua and decidualized HESC (P < 0.001, P = 0.014). Hsa_circ_0001550 is a covalently closed RNA molecule that was verified by RNase R assay and Act D assay (P = 0.012). Nuclear and cytoplasmic separation experiments confirmed that hsa_circ_0001550 was mainly distributed in the cytoplasm. Overexpression of hsa_circ_0001550 inhibited decidualization of HESC (P < 0.0001). Furthermore, overexpression of hsa_circ_0001550 inhibited proliferation by decreasing the number of S phase cells (P = 0.033). Annexin-V/propidium iodide double staining experiment and western blotting revealed that overexpression of hsa_circ_0001550 promoted HESC apoptosis (P < 0.001, P = 0.0139).

CONCLUSIONS

Hsa_circ_0001550 impairs decidualization of HESC. Progesterone decreases the expression of hsa_circ_0001550. The results may provide new insights into the cause of decidualization.

摘要

研究问题

hsa_circ_0001550 在蜕膜化中的分子功能是什么？

设计

从子宫内膜组织中分离出人子宫内膜基质细胞（HESC），建立体外蜕膜化模型。使用不同浓度的醋酸甲羟孕酮（MPA）观察 hsa_circ_0001550 的表达水平是否与孕激素有关。通过 RNase R、放线菌素 D（Act D）测定和细胞质/核质分馏测定来确定 hsa_circ_0001550 的生物学特性和分布。然后通过腺病毒载体过表达 hsa_circ_0001550。通过细胞计数试剂盒-8（CCK-8）测定细胞增殖。通过流式细胞术分析评估细胞周期。通过 Annexin-V/碘化丙啶双染实验和 Western blot 测定细胞凋亡。

结果

hsa_circ_0001550 在蜕膜和蜕膜化的 HESC 中表达下调（P < 0.001，P=0.014）。hsa_circ_0001550 是一种共价闭合的 RNA 分子，这通过 RNase R 测定和 Act D 测定得到证实（P=0.012）。核质分离实验证实 hsa_circ_0001550 主要分布在细胞质中。hsa_circ_0001550 的过表达抑制 HESC 的蜕膜化（P < 0.0001）。此外，通过减少 S 期细胞数量抑制增殖（P=0.033）。Annexin-V/碘化丙啶双染实验和 Western blot 揭示 hsa_circ_0001550 的过表达促进 HESC 凋亡（P < 0.001，P=0.0139）。