Huang YuHao, Zeng XinYu, Cai YanLing, Yang Yan, Zhang YuJie, Ma YiQi, Li SuPing
Department of Nuclear Medicine, The Affiliated Hospital of North Sichuan Medicine College, Nanchong City, Sichuan Province, China.
Turk J Biol. 2023 Dec 7;48(2):142-152. doi: 10.55730/1300-0152.2689. eCollection 2024.
BACKGROUND/AIM: Papillary thyroid carcinoma (PTC) is the most common form of thyroid cancer. The critical importance of circular RNA (circRNA) in a range of cancer types has been lately recognized. However, research on the functions of circRNAs in PTC has been limited thus far. Therefore, this research aimed at exploring the function and mechanism of circ-methyltransferase-like 15 (METTL15) in PTC cells.
Quantitative measurements of circ-METTL15, miR-200c-3p, and X-linked inhibitor of apoptosis protein (XIAP) in PTC cells were conducted using reverse transcription-quantitative polymerase chain reaction or Western blot analysis. To investigate cell growth, cell counting kit-8 and colony formation tests were employed, apoptosis was analyzed using flow cytometry, and migration and invasion were studied through Transwell assays. The targeted binding sites between miR-200c-3p and circ-METTL15 or XIAP were predicted by starBase and then verified by dual luciferase reporter assay.
circ-METTL15 and XIAP were upregulated in the PTC cells, while miR-200c-3p was downregulated. Downregulating circ-METTL15 or upregulating miR-200c-3p resulted in inhibited proliferation, migration, and invasion of PTC cells, while promoting apoptosis. miR-200c-3p was the downstream molecule of circ-METTL15, and XIAP was the direct target of miR-200c-3p. Forcing XIAP expression obstructed circ-METTL15 silencing to inhibit PTC cell activity.
By coopting miR-200c-3p/XIAP, Circ-METTL15 stimulates aggressive behavior in PTC cells.
背景/目的:甲状腺乳头状癌(PTC)是甲状腺癌最常见的形式。环状RNA(circRNA)在一系列癌症类型中的关键重要性最近已得到认可。然而,迄今为止,关于circRNA在PTC中的功能研究有限。因此,本研究旨在探讨环状甲基转移酶样15(METTL15)在PTC细胞中的功能及机制。
采用逆转录-定量聚合酶链反应或蛋白质免疫印迹分析对PTC细胞中的circ-METTL15、miR-200c-3p和X连锁凋亡抑制蛋白(XIAP)进行定量检测。为研究细胞生长,采用细胞计数试剂盒-8和集落形成试验,通过流式细胞术分析细胞凋亡,并通过Transwell实验研究细胞迁移和侵袭。通过starBase预测miR-200c-3p与circ-METTL15或XIAP之间的靶向结合位点,然后通过双荧光素酶报告基因检测进行验证。
circ-METTL15和XIAP在PTC细胞中上调,而miR-200c-3p下调。下调circ-METTL15或上调miR-200c-3p可导致PTC细胞增殖、迁移和侵袭受到抑制,同时促进细胞凋亡。miR-200c-3p是circ-METTL15的下游分子,XIAP是miR-200c-3p的直接靶标。强制表达XIAP可阻碍circ-METTL15沉默对PTC细胞活性的抑制作用。
Circ-METTL15通过利用miR-200c-3p/XIAP刺激PTC细胞的侵袭性行为。