Circ_0001971 通过在体外和体内靶向 miR-194/miR-204 调节口腔鳞状细胞癌的进展和化疗敏感性。
Circ_0001971 regulates oral squamous cell carcinoma progression and chemosensitivity by targeting miR-194/miR-204 in vitro and in vivo.
机构信息
Department of Endodontics, School and Hospital of Stomatology, Guizhou Medical University/Guizhou Medical University Affiliated Stomatological Hospital, Guiyang, Guizhou, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Mar;24(5):2470-2481. doi: 10.26355/eurrev_202003_20515.
OBJECTIVE
Circular RNAs (circRNAs) play a wide role in human cancers, including oral squamous cell carcinoma (OSCC). The purpose of this study was to investigate the biological functions of circ_0001971 and associated mechanisms in OSCC.
MATERIALS AND METHODS
The expression of circ_0001971, miR-194, and miR-204 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Cell proliferation and viability were assessed using cell counting kit-8 (CCK-8) assay. Cell migration and invasion were examined using the transwell assay. Cell apoptosis was monitored by flow cytometry assay. The protein levels of proliferation marker (CyclinD1), epithelial mesenchymal-transition (EMT) markers (E-cadherin (E-cad) and N-cadherin (N-cad)) and apoptosis markers (Cleaved-caspase-3 (Cleaved-cas-3) and Cleaved-caspase-9 (Cleaved-cas-9)) were measured by Western blot. The relationship between circ_0001971 and miR-194 or miR-204 was predicted by online tool starBase and verified by the Dual-Luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Tumor formation assay in nude mice was conducted to observe the role of circ_0001971 in vivo.
RESULTS
The expression of circ_0001971 was significantly increased in tumor tissues and cell lines. Circ_0001971 knockdown inhibited cell proliferation, migration, and invasion but promoted cisplatin (DDP) sensitivity and cell apoptosis. It was confirmed that miR-194 and miR-204 were targets of circ_0001971, and miR-194 inhibition or miR-204 inhibition reversed the effects of circ_0001971 knockdown in OSCC cells. Moreover, circ_0001971 knockdown impeded tumorigenesis and development in vivo.
CONCLUSIONS
Circ_0001971 regulates cell proliferation, migration, invasion, apoptosis, and chemosensitivity of OSCC by interacting with miR-194 and miR-204 in vitro and in vivo. We provided a theoretical basis for the action mechanism of circ_0001971 on OSCC progression and chemosensitivity.
目的
环状 RNA（circRNAs）在人类癌症中发挥广泛作用，包括口腔鳞状细胞癌（OSCC）。本研究旨在探讨 circ_0001971 在 OSCC 中的生物学功能及其相关机制。
材料与方法
采用实时定量聚合酶链反应（qRT-PCR）检测 circ_0001971、miR-194 和 miR-204 的表达。用细胞计数试剂盒-8（CCK-8）检测细胞增殖和活力。通过 Transwell 检测细胞迁移和侵袭。用流式细胞术检测细胞凋亡。Western blot 检测增殖标志物（CyclinD1）、上皮间质转化（EMT）标志物（E-钙黏蛋白（E-cad）和 N-钙黏蛋白（N-cad））和凋亡标志物（Cleaved-caspase-3（Cleaved-cas-3）和 Cleaved-caspase-9（Cleaved-cas-9））的蛋白水平。通过在线工具 starBase 预测 circ_0001971 与 miR-194 或 miR-204 的关系，并通过双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）检测进行验证。裸鼠肿瘤形成实验观察 circ_0001971 在体内的作用。
结果
circ_0001971 在肿瘤组织和细胞系中表达明显增加。circ_0001971 敲低抑制细胞增殖、迁移和侵袭，但促进顺铂（DDP）敏感性和细胞凋亡。证实 miR-194 和 miR-204 是 circ_0001971 的靶标，circ_0001971 敲低在 OSCC 细胞中逆转 miR-194 或 miR-204 的抑制作用。此外，circ_0001971 敲低抑制体内肿瘤的发生和发展。
结论
circ_0001971 通过体内外与 miR-194 和 miR-204 相互作用调节 OSCC 细胞的增殖、迁移、侵袭、凋亡和化疗敏感性。为 circ_0001971 对 OSCC 进展和化疗敏感性的作用机制提供了理论依据。
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