The First Department of General Surgery, Affiliated Dongguan People's Hospital, Southern Medical University(Dongguan People's Hospital), Dongguan, Guangdong, China.
Department of Anesthesiolopy, Affiliated Dongguan People's Hospital, Southern Medical University(Dongguan People's Hospital), Dongguan, Guangdong, China.
Hormones (Athens). 2023 Sep;22(3):375-387. doi: 10.1007/s42000-023-00461-7. Epub 2023 Jun 28.
This research aimed to explore the regulatory molecular mechanism among circular RNA (circ)_0011373, microRNA (miR)-1271, and lipoprotein receptor-related protein 6 (LRP6) in papillary thyroid carcinoma (PTC).
Quantitative real-time PCR (qRT-PCR) assay was adopted to measure the expression of circ_0011373, miR-1271, and LRP6 mRNA. Furthermore, cell cycle distribution, apoptosis, migration and invasion were investigated by flow cytometry and transwell assay, respectively. The target relationship between miR-1271 and circ_0011373 or LRP6 was predicted by using the Starbase website and DIANA TOOL and verified by dual-luciferase reporter and RIP assay. Protein expression levels of LRP6, p-mTOR, mTOR, p-AKT, AKT, p-PI3K, and PI3K were tested by Western blot. The function of circ_0011373 on PTC tumor growth was validated by the xenograft tumor model in vivo.
Circ_0011373 and LRP6 were upregulated, while miR-1271 was downregulated in PTC tissues and cell lines. Moreover, knockdown of circ_0011373 inhibited cell cycle, migration, and invasion and promoted apoptosis. Of particular importance was the fact that circ_0011373 directly interacted with miR-1271 and miR-1271 inhibitor was able to reverse the effect of circ_0011373 knockdown on PTC cell progression. Meanwhile, LRP6 was directly targeted by miR-1271, and its expression was positively regulated by circ_0011373. We further confirmed that miR-1271 overexpression suppressed cell cycle, migration, and invasion and enhanced apoptosis by regulating LRP6. In addition, circ_0011373 knockdown restrained PTC tumor growth in vivo.
Circ_0011373 might be able to regulate PTC cell cycle, migration, invasion, and apoptosis by regulating the miR-1271/LRP6 axis.
本研究旨在探讨环状 RNA（circ）_0011373、微小 RNA（miR）-1271 和脂蛋白受体相关蛋白 6（LRP6）在甲状腺乳头状癌（PTC）中的调控分子机制。
采用实时定量 PCR（qRT-PCR）检测 circ_0011373、miR-1271 和 LRP6 mRNA 的表达。此外，通过流式细胞术和 Transwell 实验分别检测细胞周期分布、凋亡、迁移和侵袭。通过 Starbase 网站和 DIANA TOOL 预测 miR-1271 与 circ_0011373 或 LRP6 的靶关系，并通过双荧光素酶报告和 RIP 实验验证。通过 Western blot 检测 LRP6、p-mTOR、mTOR、p-AKT、AKT、p-PI3K 和 PI3K 的蛋白表达水平。通过体内异种移植肿瘤模型验证 circ_0011373 对 PTC 肿瘤生长的作用。
circ_0011373 和 LRP6 在 PTC 组织和细胞系中上调，而 miR-1271 下调。此外，circ_0011373 的敲低抑制了细胞周期、迁移和侵袭，促进了凋亡。特别重要的是，circ_0011373 直接与 miR-1271 相互作用，miR-1271 抑制剂能够逆转 circ_0011373 敲低对 PTC 细胞进展的影响。同时，LRP6 是 miR-1271 的直接靶点，其表达受 circ_0011373 的正向调节。我们进一步证实，miR-1271 通过调节 LRP6 抑制细胞周期、迁移和侵袭，增强凋亡。此外，circ_0011373 的敲低抑制体内 PTC 肿瘤生长。
circ_0011373 可能通过调节 miR-1271/LRP6 轴来调节 PTC 细胞周期、迁移、侵袭和凋亡。
