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Hsa_circ_0000069 通过海绵吸附 miR-1270 促进 CPEB4 表达来加速宫颈癌进展。

Hsa_circ_0000069 Accelerates Cervical Cancer Progression by Sponging miR-1270 to Facilitate CPEB4 Expression.

机构信息

Obstetrics and Gynecology Department, Chongming Hospital Affiliated to Shanghai University of Medicine and Health Sciences, Shanghai, China.

Obstetrics and Gynecology Department, Hangzhou TCM Hospital Affiliated to Zhejiang Chinese Medical University, No. 453, Tiyuchang Road, Hangzhou, 310007, China.

出版信息

Biochem Genet. 2024 Jun;62(3):1638-1656. doi: 10.1007/s10528-023-10494-7. Epub 2023 Sep 4.

Abstract

The critical importance of circular RNAs (circRNAs) in human cancers, including cervical cancer (CC), has been discovered in recent years. However, the function and mechanism of hsa_circ_0000069 (circ_0000069) in CC have been fully understood. The expression levels of circ_0000069, microRNAs (miR-1270, miR-1276 and miR-620) and cytoplasmic polyadenylation element binding protein 4 (CPEB4) mRNA were detected by quantitative real time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, wound healing, transwell and tube formation assays were used to clarify the effects of circ_0000069 on the functional behaviors of CC cells. The binding relationships among miR-1270, circ_0000069 and CPEB4 were detected by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. A xenograft tumor model was established to explore the effect of circ_0000069 on tumor growth in vivo. Circ_0000069 was upregulated in CC clinical samples and cell lines, and its expression was associated with the clinical stage of CC patients. Circ_0000069 knockdown significantly decreased cell proliferation, invasion, migration, and tube formation and increased cell apoptosis in vitro. Moreover, miR-1270 was a direct target of circ_0000069, and CPEB4 was the downstream target of miR-1270. Knockdown of miR-1270 reversed the inhibitory effect of circ_0000069 knockdown on CC progression, and CPEB4 overexpression overturned the effect of miR-1270 on CC progression. In xenograft experiments, the oncogenic effect of circ_0000069 on tumor growth was verified. Altogether, circ_0000069 adsorbed miR-1270 to upregulate CPEB4 expression, thereby promoting the malignant phenotypes of CC cells. Circ_0000069 might be a potential target for treatment of CC.

摘要

近年来,人们发现环状 RNA(circRNA)在包括宫颈癌(CC)在内的人类癌症中的重要性。然而,hsa_circ_0000069(circ_0000069)在 CC 中的功能和机制尚未完全阐明。通过实时定量聚合酶链反应(qRT-PCR)检测 circ_0000069、microRNAs(miR-1270、miR-1276 和 miR-620)和细胞质多聚腺苷酸化元件结合蛋白 4(CPEB4)mRNA 的表达水平。使用细胞计数试剂盒-8(CCK-8)、5-乙炔基-2'-脱氧尿苷(EdU)、流式细胞术、划痕愈合、transwell 和管形成测定来阐明 circ_0000069 对 CC 细胞功能行为的影响。通过双荧光素酶报告和 RNA 免疫沉淀(RIP)测定检测 miR-1270、circ_0000069 和 CPEB4 之间的结合关系。建立异种移植肿瘤模型以研究 circ_0000069 对体内肿瘤生长的影响。circ_0000069 在 CC 临床样本和细胞系中上调,其表达与 CC 患者的临床分期相关。circ_0000069 敲低显著降低了细胞增殖、侵袭、迁移和管形成能力,并增加了细胞凋亡。此外,miR-1270 是 circ_0000069 的直接靶标,CPEB4 是 miR-1270 的下游靶标。miR-1270 敲低逆转了 circ_0000069 敲低对 CC 进展的抑制作用,CPEB4 过表达推翻了 miR-1270 对 CC 进展的影响。在异种移植实验中,验证了 circ_0000069 对肿瘤生长的致癌作用。总之,circ_0000069 吸附 miR-1270 以上调 CPEB4 表达,从而促进 CC 细胞的恶性表型。circ_0000069 可能是治疗 CC 的潜在靶点。

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