环形RNA(circ)_0053277有助于结直肠癌癌症细胞生长、血管生成、转移和糖酵解。

IF 2.4 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Molecular Biotechnology Pub Date : 2024-11-01 Epub Date: 2023-11-02 DOI:10.1007/s12033-023-00936-3
Jianbin Zhuang, Weiliang Song, Minghao Li, Di Kang, Kang Cheng
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引用次数: 0

摘要

环状RNA(circRNA)已被发现在许多癌症中异常表达,包括癌症(CRC)。Circ_0053277已被发现介导CRC恶性过程,并且可能是CRC进展的关键调节因子。因此,它在CRC过程中的作用及其潜在的分子机制值得进一步研究。定量实时PCR用于检测circ_0053277、microRNA-520 h(miR-520 h)和己糖激酶1(HK1)的表达水平。细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷测定法、流式细胞术、伤口愈合测定法、transwell测定法和试管形成测定法用于检测CRC细胞增殖、凋亡、迁移、侵袭和血管生成。免疫印迹法检测细胞凋亡相关标志物和HK1的蛋白水平。通过双荧光素酶报告基因分析、RNA免疫沉淀分析和RNA下拉分析验证了CRC细胞中circ_0053277与miR-520h或miR-520H与HK1之间的关系。通过检测葡萄糖摄取和乳酸生成来评估细胞糖酵解。通过体内异种移植物模型评估沉默的circ_0053277对CRC肿瘤生长的影响。我们的研究发现,circ_0053277在CRC组织和细胞中的表达升高。此外,circ_0053277敲低抑制CRC细胞增殖、血管生成、迁移和侵袭,同时促进细胞凋亡。在机制方面,circ_0053277吸附miR-520小时,HK1是miR-520 h的靶标。同时,miR-520 h抑制剂逆转了circ_0053277沉默对CRC细胞进展的抑制作用,HK1过表达也逆转了miR-520小时对CRC细胞生长、血管生成和转移的抑制作用。此外,circ_0053277敲低通过调节miR-520h/HK1途径抑制CRC细胞的糖酵解。此外,circ_0053277的敲除降低了CRC肿瘤在体内的生长。Circ_0053277通过miR-520 h/HK1途径促进CRC细胞生长、血管生成、转移和糖酵解,证实Circ_00532七十七可能是CRC治疗的潜在临床靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Circular RNA (circ)_0053277 Contributes to Colorectal Cancer Cell Growth, Angiogenesis, Metastasis and Glycolysis.

Circular RNAs (circRNAs) have been found to be abnormally expressed in many cancers, including colorectal cancer (CRC). Circ_0053277 has been found to mediate CRC malignant processes and may be a key regulator for CRC progression. Therefore, its role and potential molecular mechanism in CRC process deserve further investigation. Quantitative real-time PCR was used to detect the expression levels of circ_0053277, microRNA-520 h (miR-520 h) and hexokinase 1 (HK1). Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine assay, flow cytometry, wound healing assay, transwell assay, and tube formation assay were used to detect CRC cell proliferation, apoptosis, migration, invasion, and angiogenesis. The protein levels of apoptosis-related markers and HK1 were detected by western blot. The relationship between circ_0053277 and miR-520 h or miR-520 h and HK1 in CRC cells was verified by dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA pull-down assay. Cell glycolysis was assessed by detecting glucose uptake and lactate production. The effect of silenced circ_0053277 on CRC tumor growth was evaluated by xenograft model in vivo. Our study found that circ_0053277 expression was elevated in CRC tissues and cells. Moreover, circ_0053277 knockdown suppressed CRC cell proliferation, angiogenesis, migration and invasion, while promoting apoptosis. In terms of mechanism, circ_0053277 sponged miR-520 h, and HK1 was the target of miR-520 h. Meanwhile, miR-520 h inhibitor reversed the inhibitory effect of circ_0053277 silencing on CRC cell progression, and HK1 overexpression also overturned the suppressive effect of miR-520 h on CRC cell growth, angiogenesis and metastasis. Moreover, circ_0053277 knockdown inhibited the glycolysis of CRC cells by regulating miR-520 h/HK1 pathway. In addition, knockdown of circ_0053277 reduced CRC tumor growth in vivo. Circ_0053277 promoted CRC cell growth, angiogenesis, metastasis and glycolysis by miR-520 h/HK1 pathway, confirming that circ_0053277 might be a potential clinical target for CRC treatment.

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来源期刊
Molecular Biotechnology
Molecular Biotechnology 医学-生化与分子生物学
CiteScore
4.10
自引率
3.80%
发文量
165
审稿时长
6 months
期刊介绍: Molecular Biotechnology publishes original research papers on the application of molecular biology to both basic and applied research in the field of biotechnology. Particular areas of interest include the following: stability and expression of cloned gene products, cell transformation, gene cloning systems and the production of recombinant proteins, protein purification and analysis, transgenic species, developmental biology, mutation analysis, the applications of DNA fingerprinting, RNA interference, and PCR technology, microarray technology, proteomics, mass spectrometry, bioinformatics, plant molecular biology, microbial genetics, gene probes and the diagnosis of disease, pharmaceutical and health care products, therapeutic agents, vaccines, gene targeting, gene therapy, stem cell technology and tissue engineering, antisense technology, protein engineering and enzyme technology, monoclonal antibodies, glycobiology and glycomics, and agricultural biotechnology.
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