Ma Yue, Liu Jing, Yang Zhuo, Chen Peng, Wang Dan-Bo
Department of Gynecology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital and Institute, Shenyang, Liaoning 110042, P. R. China.
J Cancer. 2022 Jan 1;13(2):541-553. doi: 10.7150/jca.61437. eCollection 2022.
Cervical cancer (CC) is the highest incidence of female malignant tumor in China. Circular RNAs (circRNAs) has been reported to affect CC progression by altering mRNA stability at the transcriptional level or binding to miRNAs to produce competitive endogenous RNA (ceRNA). For this purpose, our study was aimed to investigate the function effects and the potential regulatory mechanism of the circRNA_400029 in CC cells. The expression levels of circRNA_400029 and miR-1285-3p were detected by real time polymerase chain reaction (RT-PCR). Similarly, the mRNA and protein levels of TLN1 (Talin 1) was detected by RT-PCR and Western blot. Cell-Counting Kit-8 (CCK-8), EdU and Flow cytometry assay were used to detect cell proliferation, cell cycle and apoptosis. Then the Transwell assays were used to test cell migration and invasion. Besides this, the functional targets were confirmed by Dual luciferase reporter assays. Tumor xenograft in nude mice checked the result . To begin with, circRNA_400029 was upregulated in CC cells and tissue. Knockdown circRNA_400029 inhibited cell proliferation, migration and invasion while induced cell apoptosis. Interestingly, miR-1285-3p targeted circRNA_400029 and down-regulated of miR-1285-3p could reverse the effects of circRNA_400029 weak-expression on progression and apoptosis of CC cells. Moreover, TLN1 was up-regulated in CC cells and identified as a direct target of miR-1285-3p. Meanwhile, we found that miR-1285-3p negatively regulated the function of TLN1. Finally, the circRNA_400029/miR-1285-3p/TLN1 axis could affect tumor growth The overexpressed circRNA_400029 promoted CC proliferation, migration and invasion while deduced apoptosis by sponging miR-1285-3p to regulate TLN1. CircRNA_400029 was a potential onco-circRNA in CC, and might be a promised therapy target.
宫颈癌（CC）是中国女性恶性肿瘤中发病率最高的。据报道，环状RNA（circRNAs）通过在转录水平改变mRNA稳定性或与微小RNA（miRNAs）结合产生竞争性内源RNA（ceRNA）来影响CC的进展。为此，我们的研究旨在探讨circRNA_400029在CC细胞中的功能作用及潜在调控机制。通过实时聚合酶链反应（RT-PCR）检测circRNA_400029和miR-1285-3p的表达水平。同样，通过RT-PCR和蛋白质免疫印迹法检测张力蛋白1（Talin 1，TLN1）的mRNA和蛋白质水平。使用细胞计数试剂盒-8（CCK-8）、EdU和流式细胞术检测细胞增殖、细胞周期和凋亡。然后使用Transwell实验检测细胞迁移和侵袭。此外，通过双荧光素酶报告基因实验确认功能靶点。在裸鼠中进行肿瘤异种移植以验证结果。首先，circRNA_400029在CC细胞和组织中上调。敲低circRNA_400029可抑制细胞增殖、迁移和侵袭，同时诱导细胞凋亡。有趣的是，miR-1285-3p靶向circRNA_400029，下调miR-1285-3p可逆转circRNA_400029低表达对CC细胞进展和凋亡的影响。此外，TLN1在CC细胞中上调，并被鉴定为miR-1285-3p的直接靶点。同时，我们发现miR-1285-3p负向调节TLN1的功能。最后，circRNA_400029/miR-1285-3p/TLN1轴可影响肿瘤生长。过表达的circRNA_400029通过海绵吸附miR-1285-3p来调节TLN1，从而促进CC增殖、迁移和侵袭，同时减少凋亡。CircRNA_400029是CC中一种潜在的致癌circRNA，可能是一个有前景的治疗靶点。
