Circ_0001667 敲低通过释放 miR-4458 耗尽 NCOA3 来阻断乳腺癌的癌症进展并减弱阿霉素耐药性。
Circ_0001667 knockdown blocks cancer progression and attenuates adriamycin resistance by depleting NCOA3 via releasing miR-4458 in breast cancer.
机构信息
Department of Tumor Diagnosis and Treatment Center of Chinese and Western mtegrative Medicine, The Central Hospital of Enshi Tujia And Miao Autonomous Prefectrue, Enshi City, Hubei Province, China.
Department of Oncology, The Central Hospital of Enshi Tujia And Miao Autonomous Prefectrue, Enshi City, Hubei Province, China.
出版信息
Drug Dev Res. 2022 Feb;83(1):75-87. doi: 10.1002/ddr.21845. Epub 2021 Jul 5.
Accumulating evidence suggests that developmental chemoresistance in cancers is closely associated with the dysregulation of circular RNA transcriptions. The objective of this study is to disclose the role of circ_0001667 and provide a potential functional mechanism in breast cancer. Quantitative real-time PCR was applied for the analysis of circ_0001667, microRNA-4458 (miR-4458) and nuclear receptor coactivator 3 (NCOA3) expression. In adriamycin (ADM)-resistant cell lines, we investigated cell proliferation using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay and colony formation assay. Cell migration and cell invasion were determined by transwell assay. The protein levels of multi-drug resistance-1, matrix metalloproteinases-9, cleaved-caspase3, cleaved-caspase9 and NCOA3 were detected by western blot. ADM resistance was ascertained by IC50 value using MTT assay. Cell apoptosis was checked by flow cytometry assay. The putative relationship between miR-4458 and circ_0001667 and NCOA3 was validated by pull-down assay, dual-luciferase reporter assay or RNA Immunoprecipitation assay. Circ_0001667 knockdown inhibited MCF-7/ADM and MDA-MB-231/ADM cell proliferation, migration, invasion and ADM resistance. MiR-4458 was a target of circ_0001667, and its expression was decreased in ADM-resistant tumor tissues and cells. MiR-4458 inhibition reversed the effects of circ_0001667 knockdown. In depth, NCOA3 was a target of circ_0001667, and circ_0001667 knockdown weakened NCOA3 expression by releasing miR-4458. MCF-7/ADM and MDA-MB-231/ADM cell proliferation, migration, invasion, and ADM resistance inhibited by miR-4458 restoration were recovered by NCOA3 overexpression. Circ_0001667 knockdown also repressed tumor growth and ADM resistance in vivo. Circ_0001667 knockdown blocks cancer progression and attenuates ADM resistance by depleting NCOA3 via releasing miR-4458 in breast cancer.
越来越多的证据表明，癌症发育性化疗耐药性与环状 RNA 转录的失调密切相关。本研究的目的是揭示 circ_0001667 的作用，并为乳腺癌提供一个潜在的功能机制。采用实时定量 PCR 分析 circ_0001667、微小 RNA-4458 (miR-4458) 和核受体共激活因子 3 (NCOA3) 的表达。在阿霉素 (ADM) 耐药细胞系中，我们通过 3-[4,5-二甲基噻唑-2-基]-2,5 二苯基四氮唑溴盐 (MTT) 试验和集落形成试验检测细胞增殖。通过 Transwell 试验测定细胞迁移和细胞侵袭。通过蛋白质印迹法检测多药耐药蛋白 1、基质金属蛋白酶 9、裂解 caspase3、裂解 caspase9 和 NCOA3 的蛋白水平。通过 MTT 试验测定 ADM 耐药性的 IC50 值。通过流式细胞术检测细胞凋亡。通过下拉试验、双荧光素酶报告基因试验或 RNA 免疫沉淀试验验证 miR-4458 与 circ_0001667 和 NCOA3 之间的假定关系。circ_0001667 敲低抑制 MCF-7/ADM 和 MDA-MB-231/ADM 细胞增殖、迁移、侵袭和 ADM 耐药性。miR-4458 是 circ_0001667 的靶标，其在 ADM 耐药性肿瘤组织和细胞中表达降低。miR-4458 抑制逆转了 circ_0001667 敲低的作用。深入研究表明，NCOA3 是 circ_0001667 的靶标，circ_0001667 敲低通过释放 miR-4458 减弱 NCOA3 的表达。miR-4458 恢复抑制的 MCF-7/ADM 和 MDA-MB-231/ADM 细胞增殖、迁移、侵袭和 ADM 耐药性被 NCOA3 过表达所恢复。circ_0001667 敲低也抑制了体内肿瘤生长和 ADM 耐药性。circ_0001667 敲低通过释放 miR-4458 耗尽 NCOA3 从而抑制乳腺癌的肿瘤进展并减轻 ADM 耐药性。
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