Zhang Jun, Zhang Yan, Li Xing, Bao Yindi, Yang Jing
Department of Obstetrics and Gynecology, Renmin Hospital of Wuhan University, Wuhan, China.
Reproductive Medical Center/Hubei Medical Clinical Research Center for Assisted Reproductive Technology and Embryonic Development, Renmin Hospital of Wuhan University, Wuhan, China.
Cancer Gene Ther. 2025 Feb;32(2):214-226. doi: 10.1038/s41417-024-00866-5. Epub 2025 Jan 2.
Cervical cancer (CC) is a prevalent gynecological malignancy. Increasing evidence suggests that circular RNAs (circRNAs) play a pivotal role in the pathogenesis of CC. However, the regulatory function of circ_ASH1L in CC remains elusive. In this study, we aim to elucidate the precise role and underlying mechanism of circ_ASH1L in the malignant progression of CC. The human CC dataset GSE102686 was extracted from the Gene Expression Omnibus (GEO) database for the analysis of differentially expressed circRNAs. Target gene prediction softwares were utilized to predict the binding of miRNAs to circ_ASH1L sponge. The expression level of circ_ASH1L in CC tissues and cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The characteristics of circ_ASH1L were determined by RNase R digestion, actinomycin D, and nucleo-plasmic separation assays. The effects of circ_ASH1L, miR-1254, and CD36 gain-and-loss on the malignant progression of CC were investigated using Cell Counting Kit-8 (CCK-8), colony formation, flow cytometry, wound scratch, transwell, and Western blot assay. The effect of circ_ASH1L on tumorigenicity of CC cells in vivo was evaluated in nude mice through tumor xenograft assay. The targeted regulatory relationship between circ_ASH1L/miR-1254 as well as miR-1254/CD36 was validated by dual-luciferase reporter assay. We screened the differentially expressed circ_ASH1L from the GEO dataset GSE102686 and confirmed its circular structure. Furthermore, we observed a significant upregulation of circ_ASH1L in both CC tissues and cells. Overexpression of circ_ASH1L promotes proliferation, invasion, and migration of CC cells while inhibiting cell apoptosis. However, silencing circ_ASH1L showed opposite results and inhibited tumorigenicity of CC cells in nude mice. Furthermore, we have identified circ_ASH1L as a miR-1254 sponge in CC cells. Notably, our in vitro experiments demonstrated that exogenously modulating the expression of miR-1254 effectively counteracted the impact of circ_ASH1L on the malignant phenotypic characteristics of CC cells. Similarly, modulation of CD36 expression efficiently counteracted the effect of miR-1254 on the malignant biological behavior of CC cells. In conclusion, circ_ASH1L promoted the malignant progression of CC via upregulating CD36 expression through sponging miR-1254.
宫颈癌（CC）是一种常见的妇科恶性肿瘤。越来越多的证据表明，环状RNA（circRNAs）在CC的发病机制中起关键作用。然而，circ_ASH1L在CC中的调控功能仍不清楚。在本研究中，我们旨在阐明circ_ASH1L在CC恶性进展中的精确作用及潜在机制。从基因表达综合数据库（GEO）中提取人类CC数据集GSE102686，用于分析差异表达的circRNAs。利用靶基因预测软件预测miRNAs与circ_ASH1L海绵的结合。通过定量实时聚合酶链反应（qRT-PCR）检测CC组织和细胞中circ_ASH1L的表达水平。通过RNase R消化、放线菌素D和核质分离试验确定circ_ASH1L的特征。使用细胞计数试剂盒-8（CCK-8）、集落形成、流式细胞术、伤口划痕、Transwell和蛋白质印迹分析研究circ_ASH1L、miR-1254和CD36的增减对CC恶性进展 的影响。通过肿瘤异种移植试验在裸鼠中评估circ_ASH1L对CC细胞体内致瘤性的影响。通过双荧光素酶报告基因试验验证circ_ASH1L/miR-1254以及miR-1254/CD36之间的靶向调控关系。我们从GEO数据集GSE102686中筛选出差异表达的circ_ASH1L，并证实了其环状结构。此外，我们观察到circ_ASH1L在CC组织和细胞中均显著上调。circ_ASH1L的过表达促进CC细胞的增殖、侵袭和迁移，同时抑制细胞凋亡。然而，沉默circ_ASH1L则显示出相反的结果，并抑制了裸鼠中CC细胞的致瘤性。此外，我们已确定circ_ASH1L是CC细胞中的miR-1254海绵。值得注意的是，我们的体外实验表明，外源调节miR-1254的表达有效地抵消了circ_ASH1L对CC细胞恶性表型特征的影响。同样，调节CD36表达有效地抵消了miR-1254对CC细胞恶性生物学行为的影响。总之，circ_ASH1L通过充当miR-1254的海绵上调CD36表达，从而促进CC的恶性进展。
