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微小RNA-320b通过靶向c-Myc抑制人结肠癌细胞的增殖。

miR-320b suppresses cell proliferation by targeting c-Myc in human colorectal cancer cells.

作者信息

Wang Hantao, Cao Fuao, Li Xu, Miao Hua, E Jifu, Xing Junjie, Fu Chuan-Gang

机构信息

Department of Colorectal Surgery, Changhai Hospital, Shanghai, 200433, China.

Department of General Surgery, The First People's Hospital of Pinghu, Pinghu, 314200, Zhejiang Province, China.

出版信息

BMC Cancer. 2015 Oct 20;15:748. doi: 10.1186/s12885-015-1728-5.

DOI:10.1186/s12885-015-1728-5
PMID:26487644
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4617986/
Abstract

BACKGROUND

MicroRNAs (miRNAs) are small noncoding RNAs that potentially play a critical role in tumorigenesis. Mounting evidence indicates that one specific miRNA: miR-320b is down regulated in numerous human cancers, including colorectal cancer (CRC); making the hypothesis that miR-320b may play a key role in tumorigenesis plausible. However, its role in carcinogenesis remains poorly defined. The goal of this study is to better clarify the role of miR-320b in tumor growth of CRC.

METHODS

Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was conducted to detect the expression of miR-320b in CRC tissues and 5 CRC cell lines. The effect of miR-320b on cell proliferation was analyzed in vitro and in vivo. Furthermore, a luciferase reporter assay was performed to measure the target effects of miR-320b. Lastly, the messenger RNA (mRNA) and protein levels of the gene c-MYC were measured in CRC cell lines and tissues by qRT-PCR, and confirmed via Western blot and Immunohistochemical (IHC) staining.

RESULTS

The results presented here showed that miR-320b expression was down regulated in both CRC tissues and cells. Overexpression of miR-320b in CRC cells was statistically correlated with a decrease of cell growth in vitro and in vivo, while c-MYC was identified as a target gene of miR-320b in CRC. Furthermore, it was found that up-regulation of c-Myc can attenuate the effects induced by miR-320b.

CONCLUSIONS

Our identification of c-MYC as a target gene of miR-320b provides new insights into the pathophysiology of CRC proliferation, and identifies miR-320b as a novel therapeutic target for the treatment of CRC.

摘要

背景

微小RNA(miRNA)是一类小的非编码RNA,可能在肿瘤发生过程中发挥关键作用。越来越多的证据表明,一种特定的miRNA:miR-320b在包括结直肠癌(CRC)在内的多种人类癌症中表达下调;这使得miR-320b可能在肿瘤发生中起关键作用这一假设变得合理。然而,其在致癌过程中的作用仍不清楚。本研究的目的是更好地阐明miR-320b在CRC肿瘤生长中的作用。

方法

采用定量逆转录聚合酶链反应(qRT-PCR)检测CRC组织和5种CRC细胞系中miR-320b的表达。在体外和体内分析了miR-320b对细胞增殖的影响。此外,进行了荧光素酶报告基因检测以测量miR-320b的靶向作用。最后,通过qRT-PCR测量CRC细胞系和组织中c-MYC基因的信使RNA(mRNA)和蛋白质水平,并通过蛋白质印迹法和免疫组织化学(IHC)染色进行确认。

结果

此处呈现的结果表明,miR-320b在CRC组织和细胞中均表达下调。CRC细胞中miR-320b的过表达在统计学上与体外和体内细胞生长的减少相关,而c-MYC被鉴定为CRC中miR-320b的靶基因。此外,发现c-Myc的上调可减弱miR-320b诱导的作用。

结论

我们将c-MYC鉴定为miR-320b的靶基因为CRC增殖的病理生理学提供了新的见解,并将miR-320b鉴定为治疗CRC的新治疗靶点。

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