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Hsa_circ_0137008通过充当微小RNA-338-5p的海绵来抑制结直肠癌的恶性表型。

Hsa_circ_0137008 suppresses the malignant phenotype in colorectal cancer by acting as a microRNA-338-5p sponge.

作者信息

Yang Zhanfeng, Zhang Jingjing, Lu Danghui, Sun Yan, Zhao Xinyong, Wang Xiaoqiong, Zhou Wen, He Qunli, Jiang Zhi

机构信息

1Department of Medicine, Zhengzhou University of Industry Technology, 16 Xueyuan Road, Xinzheng, 451100 Henan China.

2Department of Vascular Surgery, Henan Provincial People's Hospital, 7 Weiwu Road, Zhengzhou, 450000 Henan China.

出版信息

Cancer Cell Int. 2020 Mar 4;20:67. doi: 10.1186/s12935-020-1150-1. eCollection 2020.

Abstract

BACKGROUND

Circular RNAs (circRNAs) have been shown to play a crucial role in tumorigenesis. In this study, we investigated the function of hsa_circ_0137008 and its underlying molecular mechanism in colorectal cancer (CRC).

METHODS

Gene expression was conducted by quantitative real-time PCR or western blot. Functional experiments were performed by cell count kit-8, colony formation assay, wound healing, and transwell assays. Luciferase reporter assay and RNA pull-down assay were performed to investigate the molecular mechanism of hsa_circ_0137008 in CRC. In addition, the xenograft tumor model was applied to determine the role of hsa_circ_0137008 in vivo.

RESULTS

Downregulation of hsa_circ_0137008 was observed in CRC tissues and cell lines. Functionally, overexpression of hsa_circ_0137008 inhibited the proliferation of CRC cells, as indicated by the inhibition of proliferative protein expression (Ki67 and PCNA), reduced cell viability and colony formation ability. Upregulation of hsa_circ_0137008 suppressed the migration, invasion, and epithelial to mesenchymal transition (EMT) of CRC cells. Mechanically, hsa_circ_0137008 negatively regulated the expression of microRNA-338-5p (miR-338-5p). Furthermore, hsa_circ_0137008 abated the miR-338-5p mediated promotion on CRC cell progression. Tumor suppressive function of hsa_circ_0137008 was validated in vivo.

CONCLUSION

These findings highlighted the fact that overexpression of hsa_circ_0137008 inhibited the progression of CRC via sponging miR-338-5p, suggesting that hsa_circ_0137008/miR-338-5p axis is a principal regulator of CRC tumorigenesis.

摘要

背景

环状RNA(circRNAs)已被证明在肿瘤发生中起关键作用。在本研究中,我们调查了hsa_circ_0137008在结直肠癌(CRC)中的功能及其潜在分子机制。

方法

通过定量实时PCR或蛋白质免疫印迹法检测基因表达。通过细胞计数试剂盒-8、集落形成试验、伤口愈合试验和Transwell试验进行功能实验。进行荧光素酶报告基因试验和RNA下拉试验以研究hsa_circ_0137008在CRC中的分子机制。此外,应用异种移植肿瘤模型确定hsa_circ_0137008在体内的作用。

结果

在CRC组织和细胞系中观察到hsa_circ_0137008表达下调。在功能上,hsa_circ_0137008的过表达抑制了CRC细胞的增殖,这表现为增殖蛋白表达(Ki67和PCNA)受到抑制、细胞活力降低以及集落形成能力下降。hsa_circ_0137008的上调抑制了CRC细胞的迁移、侵袭以及上皮-间质转化(EMT)。从机制上讲,hsa_circ_0137008负向调节微小RNA-338-(miR-338-5p)的表达。此外,hsa_circ_0137008减弱了miR-338-5p介导的对CRC细胞进展的促进作用。hsa_circ_0137008的肿瘤抑制功能在体内得到验证。

结论

这些发现突出了以下事实,即hsa_circ_0137008的过表达通过海绵吸附miR-338-5p抑制了CRC的进展,表明hsa_circ_0137008/miR-338-5p轴是CRC肿瘤发生的主要调节因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/176f/7057602/8134edb03461/12935_2020_1150_Fig1_HTML.jpg

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