Department of Radiation Oncology, the First Affiliated Hospital of Anhui Medical University, No.218, Jixi Road, Hefei, 230022, Anhui, China.
Hormones (Athens). 2022 Dec;21(4):611-623. doi: 10.1007/s42000-022-00378-7. Epub 2022 Jul 8.
This study aimed to explore the role and underlying mechanism of circular RNA (circRNA) reticulon 1 (circRTN1) in thyroid cancer (TC).
The expression levels of circRTN1, microRNA-431-5p (miR-431-5p), and transforming growth factor-alpha (TGFA) mRNA were measured by quantitative real-time PCR (qRT-PCR). Cell proliferation was evaluated using colony formation and 5-ethynyl-2'-deoxyuridine (EdU) assays. Cell apoptosis was analyzed using flow cytometry. Cell migration and invasion were measured using the transwell assay. The protein levels of ki-67, Bax, matrix metalloproteinase 2 (MMP-2), and TGFA were detected using Western blot assay. The interaction between miR-431-5p and circRTN1 or TGFA was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. The effect of circRTN1on TC in vivo was explored via xenograft tumor assay.
The expression of circRTN1 was increased in TC tissues and cells. Knockdown of circRTN1 suppressed TC cell proliferation, migration, and invasion, and increased cell apoptosis. MiR-431-5p was a target of circRTN1, and miR-431-5p downregulation reversed the role of circRTN1 knockdown in TC cells. TGFA was identified as a direct target of miR-431-5p, and miR-431-5p exerted the anti-tumor role in TC cells by downregulating TGFA. Moreover, circRTN1 sponged miR-431-5p to regulate TGFA expression. Furthermore, circRTN1 knockdown inhibited tumor growth in vivo.
CircRTN1 acted as a cancer-promoting circRNA in TC by regulating the miR-431-5p/TGFA axis, providing a potential therapeutic strategy for TC treatment.
本研究旨在探讨环状 RNA（circRNA）雷杜丁 1（circRTN1）在甲状腺癌（TC）中的作用及其潜在机制。
采用实时定量 PCR（qRT-PCR）检测 circRTN1、微小 RNA-431-5p（miR-431-5p）和转化生长因子-α（TGFA）mRNA 的表达水平。通过集落形成和 5-乙炔基-2'-脱氧尿苷（EdU）实验评估细胞增殖。采用流式细胞术分析细胞凋亡。通过 Transwell 实验检测细胞迁移和侵袭。采用 Western blot 实验检测 ki-67、Bax、基质金属蛋白酶 2（MMP-2）和 TGFA 的蛋白水平。通过双荧光素酶报告基因实验和 RNA 免疫沉淀（RIP）实验验证 miR-431-5p 与 circRTN1 或 TGFA 的相互作用。通过异种移植肿瘤实验探讨 circRTN1 对 TC 的体内作用。
circRTN1 在 TC 组织和细胞中表达上调。circRTN1 敲低抑制 TC 细胞增殖、迁移和侵袭，增加细胞凋亡。miR-431-5p 是 circRTN1 的靶基因，miR-431-5p 下调逆转了 circRTN1 敲低对 TC 细胞的作用。TGFA 是 miR-431-5p 的直接靶基因，miR-431-5p 通过下调 TGFA 发挥 TC 细胞的抗肿瘤作用。此外，circRTN1 作为 miR-431-5p 的海绵分子调节 TGFA 表达。进一步的，circRTN1 敲低抑制体内肿瘤生长。
circRTN1 通过调节 miR-431-5p/TGFA 轴在 TC 中发挥促癌作用，为 TC 治疗提供了一种潜在的治疗策略。
