Department of Gynecology and Obstetrics, Dalian Municipal Women and Children's Medical Center (Group), Dalian, China.
Department of Pathology, Dalian Liaoyu Hospital, Dalian, China.
Kaohsiung J Med Sci. 2022 Mar;38(3):218-229. doi: 10.1002/kjm2.12478. Epub 2021 Nov 26.
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.
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。
