Wang Hantao, Cao Fuao, Li Xu, Miao Hua, E Jifu, Xing Junjie, Fu Chuan-Gang
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.
BMC Cancer. 2015 Oct 20;15:748. doi: 10.1186/s12885-015-1728-5.
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.
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.
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.
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.
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。
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。
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。
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。