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MiR-520a-3p在调控结直肠癌中表皮生长因子受体(EGFR)表达方面的新作用

A Novel Role for MiR-520a-3p in Regulating EGFR Expression in Colorectal Cancer.

作者信息

Zhang Rui, Liu Rui, Liu Chang, Niu Yahan, Zhang Jianguo, Guo Baoliang, Zhang Chen-Yu, Li Jing, Yang Jie, Chen Xi

机构信息

State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology, NJU Advanced Institute for Life Sciences (NAILS), School of Life Sciences, Nanjing University, Nanjing, China.

State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, China.

出版信息

Cell Physiol Biochem. 2017;42(4):1559-1574. doi: 10.1159/000479397. Epub 2017 Jul 24.

Abstract

BACKGROUND/AIMS: MicroRNAs (miRNAs) have been consistently demonstrated to be involved in colorectal cancer as either tumour oncogenes or tumour suppressors. However, the detailed role of miR-520a-3p in colorectal cancer remains poorly understood.

METHODS

Quantitative RT-PCR and western blotting assays were used to measure miR-520a-3p and EGFR expression levels in colorectal cancer tissues, respectively. Luciferase reporter assay was employed to validate the direct targeting of EGFR by miR-520a-3p. Cell migration, apoptosis and cell cycle assays were performed to analyse the biological functions of miR-520a-3p and EGFR in colorectal cancer cells. In vivo experiment was performed to analyse the effects of miR-520a-3p and EGFR on the growth of colorectal cancer xenografts in mice.

RESULTS

In this study, we found that miR-520a-3p was most likely to target the EGFR 3'-UTR, which was experimentally validated. In addition, we investigated the biological effects of EGFR inhibition by miR-520a-3p both in vitro and In vivo and found that miR-520a-3p could suppress cell migration, promote apoptosis, lead to colorectal cancer cell cycle arrest at the G0/G1 phase, and decelerate tumour growth in xenograft mice, potentially by targeting EGFR.

CONCLUSIONS

This study highlights a tumour suppressor role for miR-520a-3p in colorectal cancer via the regulation of EGFR expression. Thus, miR-520a-3p may be a novel molecular therapeutic target for colorectal cancer.

摘要

背景/目的:微小RNA(miRNA)一直被证明在结直肠癌中作为肿瘤癌基因或肿瘤抑制因子发挥作用。然而,miR-520a-3p在结直肠癌中的具体作用仍知之甚少。

方法

分别采用定量逆转录-聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法检测结直肠癌组织中miR-520a-3p和表皮生长因子受体(EGFR)的表达水平。采用荧光素酶报告基因检测法验证miR-520a-3p对EGFR的直接靶向作用。进行细胞迁移、凋亡和细胞周期检测,以分析miR-520a-3p和EGFR在结直肠癌细胞中的生物学功能。进行体内实验,分析miR-520a-3p和EGFR对小鼠结直肠癌异种移植瘤生长的影响。

结果

在本研究中,我们发现miR-520a-3p最有可能靶向EGFR的3'-非翻译区(3'-UTR),这一结果得到了实验验证。此外,我们在体外和体内研究了miR-520a-3p抑制EGFR的生物学效应,发现miR-520a-3p可能通过靶向EGFR抑制细胞迁移、促进凋亡、使结直肠癌细胞周期停滞在G0/G1期,并减缓异种移植小鼠肿瘤的生长。

结论

本研究通过调节EGFR表达突出了miR-520a-3p在结直肠癌中的肿瘤抑制作用。因此,miR-520a-3p可能是结直肠癌的一个新的分子治疗靶点。

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