Department of Obstetrics and Gynaecology, Jingzhou Hospital, Yangtze University, Jingzhou, Hubei, P.R. China.
Department of Obstetrics and Gynecology, Renmin Hospital, Hubei University of Medicine, Shiyan, P.R, China.
Bioengineered. 2022 Mar;13(3):5251-5261. doi: 10.1080/21655979.2022.2037870.
Cervical cancer (CC) is one of the most common gynecological tumors worldwide. Several studies have reported that circular RNAs (circRNAs) play important roles in various types of diseases, including cancer. Thus, the present study aimed to investigate the role of circRNA_0000285 in CC development. Dual-luciferase reporter and RNA pull-down assays were performed to verify the binding region between circRNA_0000285 and miR-654-3p. The expression levels of circRNA_0000285 and miR-654-3p were analyzed in CC and the corresponding normal tissues, as well as in SiHa, HeLa, and NC104 cells using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In addition, the effect of circRNA_0000285 inhibition on cell viability, apoptosis, and the expression of apoptosis-related markers was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), flow cytometry, and Western blotting assays, respectively. The results verified that miR-654-3p directly targeted circRNA_0000285 expression. circRNA_0000285 was overexpressed and miR-654-3p expression was downregulated in CC tissues and cells compared to that in control. Moreover, circRNA_0000285 knockdown suppressed the viability and promoted the apoptosis of CC cells, which was accompanied by the downregulated and upregulated expressions B-cell lymphoma-2 () and Bcl-2 associated X (), respectively. The ratio of Bax/Bcl-2 levels also increased following circRNA_0000285 knockdown. However, these findings were abrogated after miR-654-3p inhibitor treatment. Hence, circRNA_0000285 knockdown suppressed cell viability and promoted apoptosis by targeting miR-654-3p in CC.
宫颈癌 (CC) 是全球最常见的妇科肿瘤之一。多项研究报道,环状 RNA (circRNA) 在包括癌症在内的各种疾病中发挥重要作用。因此,本研究旨在探讨 circRNA_0000285 在 CC 发展中的作用。双荧光素酶报告和 RNA 下拉实验验证了 circRNA_0000285 和 miR-654-3p 之间的结合区域。采用逆转录定量聚合酶链反应 (RT-qPCR) 分析 CC 及相应正常组织、SiHa、HeLa 和 NC104 细胞中 circRNA_0000285 和 miR-654-3p 的表达水平。此外,通过 MTT (3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)、流式细胞术和 Western blot 实验分别评估 circRNA_0000285 抑制对细胞活力、凋亡和凋亡相关标志物表达的影响。结果证实 miR-654-3p 可直接靶向 circRNA_0000285 表达。与对照相比,CC 组织和细胞中 circRNA_0000285 表达上调,miR-654-3p 表达下调。此外,circRNA_0000285 敲低抑制 CC 细胞活力并促进其凋亡,同时下调和上调 B 细胞淋巴瘤-2 () 和 Bcl-2 相关 X () 的表达。circRNA_0000285 敲低后 Bax/Bcl-2 水平比值也增加。然而,miR-654-3p 抑制剂处理后,这些发现被逆转。因此,circRNA_0000285 通过靶向 miR-654-3p 抑制 CC 细胞活力并促进其凋亡。