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环状 RNA circ_0023404 通过上调 CYP2S1 作为 miR-636 的海绵促进宫颈癌细胞的恶性行为。

Circular RNA circ_0023404 serves as a miR-636 sponge to promote malignant behaviors in cervical cancer cells through upregulation of CYP2S1.

机构信息

Department of Gynecology and Obstetrics, Dalian Municipal Women and Children's Medical Center (Group), Dalian, China.

Department of Pathology, Dalian Liaoyu Hospital, Dalian, China.

出版信息

Kaohsiung J Med Sci. 2022 Mar;38(3):218-229. doi: 10.1002/kjm2.12478. Epub 2021 Nov 26.

Abstract

Cervical cancer is the most common malignant gynecological tumor. Circular RNA (circRNA) circ_0023404 is reported to be upregulated in cervical cancer cells. This aim is to explore the role and mechanism of circ_0023404 in cervical cancer. circ_0023404, microRNA-636 (miR-636), and cytochrome P450 2S1 (CYP2S1) levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation, migration, invasion, and apoptosis were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, 5-ethynyl-2'-deoxyuridine (EDU) assay, colony formation assay, transwell assay, and cytometry assay. Protein levels of cyclin D1, matrix metallopeptidase 9 (MMP9), Bcl-2-associated X protein (Bax), and CYP2S1 were examined by western blot assay. The binding relationship between miR-636 and circ_0023404 or CYP2S1 was predicted by Circinteractome or targetscan, and then verified by a dual-luciferase reporter assay and RNA pull-down assay. circ_0023404 and CYP2S1 expression were increased, and miR-636 was decreased in cervical cancer tissues and cells. Moreover, circ_0023404 knockdown could repress proliferation, migration, invasion, and promote apoptosis of cervical cancer cells in vitro. Mechanically, circ_0023404 could regulate CYP2S1 expression by sponging miR-636. circ_0023404 silencing could attenuate the progression of cervical cancer cells partly by targeting the miR-636/CYP2S1 axis, hinting at a promising therapeutic target for cervical cancer.

摘要

宫颈癌是最常见的妇科恶性肿瘤。环状 RNA (circRNA) circ_0023404 据报道在宫颈癌细胞中上调。本研究旨在探讨 circ_0023404 在宫颈癌中的作用和机制。通过实时定量聚合酶链反应 (RT-qPCR) 检测 circ_0023404、微小 RNA-636 (miR-636) 和细胞色素 P450 2S1 (CYP2S1) 的水平。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐 (MTT) 测定法、5-乙炔基-2'-脱氧尿苷 (EDU) 测定法、集落形成测定法、Transwell 测定法和细胞术测定法检测细胞增殖、迁移、侵袭和凋亡。通过 Western blot 测定法检测细胞周期蛋白 D1、基质金属蛋白酶 9 (MMP9)、Bcl-2 相关 X 蛋白 (Bax) 和 CYP2S1 的蛋白水平。通过 Circinteractome 或 targetscan 预测 miR-636 与 circ_0023404 或 CYP2S1 的结合关系,然后通过双荧光素酶报告基因测定法和 RNA 下拉测定法进行验证。circ_0023404 和 CYP2S1 的表达在宫颈癌组织和细胞中增加,而 miR-636 的表达减少。此外,circ_0023404 敲低可抑制宫颈癌细胞在体外的增殖、迁移、侵袭并促进凋亡。机制上,circ_0023404 可通过海绵吸附 miR-636 调节 CYP2S1 的表达。circ_0023404 沉默可通过靶向 miR-636/CYP2S1 轴部分减轻宫颈癌细胞的进展,提示其为宫颈癌有前途的治疗靶点。

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