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人源环状RNA hsa_circ_0005571促进结肠癌细胞的增殖和侵袭。

Hsa_circ_0005571 promotes the proliferation and invasion of colorectal cancer cells.

作者信息

Ge Haipeng, Zhang Jiaxuan, Tao Ran, Ding Li, Jiang Song, Shen Jiale, Bian Longjun, Qin Jun

机构信息

Department of Trauma Center, Affiliated Hospital of Nantong University, Nantong City, 226000, Jiangsu Province, People's Republic of China.

Department of General Surgery, Affiliated Hospital of Nantong University, No.20 Xisi Road, Chongchuan District, Nantong City, 226000, Jiangsu Province, People's Republic of China.

出版信息

Sci Rep. 2025 Aug 22;15(1):30931. doi: 10.1038/s41598-025-16450-2.

Abstract

Dysregulated expression of circular RNAs (circRNAs) has been implicated in the initiation and progression of various diseases, including cancer. In this study, we investigated the role of hsa_circ_0005571 in modulating the biological characteristics of colorectal cancer (CRC) cells. Initially, the circRNA expression profiles of CRC tissues and corresponding normal tissues were analyzed using bioinformatics to identify differentially expressed circRNAs. Subsequently, quantitative real-time PCR (qRT‒PCR) was used to validate the expression levels of hsa_circ_0005571 in both CRC tissues and cell lines. In vitro assays, including the Cell Counting Kit-8 (CCK-8), colony formation, 5-ethynyl-2'-deoxyuridine (EdU) incorporation, wound healing, and Transwell invasion assays, were employed to assess the effects of hsa_circ_0005571 on CRC cell proliferation and invasion. Additionally, tumorigenic potential was examined through in vivo tumorigenesis experiments in nude mice. Bioinformatic predictions and experimental evidence indicated that hsa_circ_0005571 may function as a sponge for miR-520f-3p, which was found to be downregulated in CRC cells. Moreover, LRP8 was predicted and confirmed as a direct downstream target of miR-520f-3p, with both the mRNA and protein levels of LRP8 significantly elevated in CRC cells. Overall, our results suggest that hsa_circ_0005571 enhances CRC proliferation and invasion by modulating the miR-520f-3p/LRP8 axis. Furthermore, elevated hsa_circ_0005571 expression in CRC tissue samples and cell lines relative to that in normal controls was positively correlated with metastasis (p = 0.029) and advanced clinical stage (p = 0.006). Survival analyses revealed that CRC patients with high levels of hsa_circ_0005571 expression had significantly lower overall survival rates than did those with low expression (p < 0.05). Functional assays confirmed that hsa_circ_0005571 facilitates CRC cell proliferation and migration, whereas its knockdown results in reduced migration, proliferation, and tumorigenic potential in vivo. Finally, Western blot analyses demonstrated that LRP8 expression varied in accordance with the levels of hsa_circ_0005571 and miR-520f-3p, further confirming the involvement of the hsa_circ_0005571/miR-520f-3p/LRP8 regulatory axis in CRC. Our study suggested that hsa_circRNA_0005571 promotes proliferation and migration through the miR-520f-3p/LRP8 axis in CRC. Consequently, hsa_circ_0005571 may represent a promising novel gene target for the diagnosis and treatment of CRC.

摘要

环状RNA(circRNAs)表达失调与包括癌症在内的多种疾病的发生和发展有关。在本研究中,我们调查了hsa_circ_0005571在调节结直肠癌(CRC)细胞生物学特性中的作用。首先,利用生物信息学分析CRC组织和相应正常组织的circRNA表达谱,以鉴定差异表达的circRNAs。随后,采用定量实时PCR(qRT‒PCR)验证hsa_circ_0005571在CRC组织和细胞系中的表达水平。体外实验,包括细胞计数试剂盒-8(CCK-8)、集落形成、5-乙炔基-2'-脱氧尿苷(EdU)掺入、伤口愈合和Transwell侵袭实验,用于评估hsa_circ_0005571对CRC细胞增殖和侵袭的影响。此外,通过裸鼠体内肿瘤发生实验检测致瘤潜力。生物信息学预测和实验证据表明,hsa_circ_0005571可能作为miR-520f-3p的海绵,而miR-520f-3p在CRC细胞中表达下调。此外,低密度脂蛋白受体相关蛋白8(LRP8)被预测并确认为miR-520f-3p的直接下游靶点,CRC细胞中LRP8的mRNA和蛋白水平均显著升高。总体而言,我们的结果表明,hsa_circ_0005571通过调节miR-520f-3p/LRP8轴增强CRC的增殖和侵袭。此外,与正常对照相比,CRC组织样本和细胞系中hsa_circ_0005571表达升高与转移(p = 0.029)和临床晚期(p = 0.006)呈正相关。生存分析显示,hsa_circ_0005571高表达的CRC患者总生存率显著低于低表达患者(p < 0.05)。功能实验证实,hsa_circ_0005571促进CRC细胞增殖和迁移,而其敲低导致体内迁移、增殖和致瘤潜力降低。最后,蛋白质免疫印迹分析表明,LRP8表达随hsa_circ_0005571和miR-520f-3p水平而变化,进一步证实hsa_circ_0005571/miR-520f-3p/LRP8调控轴参与CRC的发生发展。我们的研究表明,hsa_circRNA_0005571通过miR-520f-3p/LRP8轴促进CRC的增殖和迁移。因此,可以认为hsa_circ_0005571是CRC诊断和治疗中有前景的新基因靶点。

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