Lu Lei, Cai Ming, Peng Meixia, Wang Fei, Zhai Xiaofeng
Department of Gastrointestinal Surgery, The Second Affiliated Hospital of Nantong University, Nantong, Jiangsu, China,
Cancer Manag Res. 2019 Feb 22;11:1805-1816. doi: 10.2147/CMAR.S183085. eCollection 2019.
Dysregulation of miRNAs is critically implicated in tumorigenesis, and aberrant expression of miR-491-5p has been reported to play a key role in initiation and progression of various cancers. However, the biological function and underlying mechanism of miR-491-5p in colorectal cancer (CRC) remain elusive.
Quantitative real-time PCR (qRT-PCR) was employed to evaluate the levels of miR-491-5p and IGF2 mRNA expression in CRC tissues, cell lines and plasma. Cell counting kit-8 and colony formation assays were used to detect the effects of miR-491-5p on CRC cell growth. Luciferase reporter assays were applied to confirm the miR-491-5p target gene. In vivo experiments were conducted in nude mice.
miR-491-5p was found to be obviously downregulated in CRC tissues and cell lines, and decreased miR-491-5p expression level was shown to be associated with differentiation, TNM stage and poor overall survival (OS). miR-491-5p overexpression suppressed CRC cell proliferation both in vitro and in vivo. Mechanically, insulin-like growth factor 2 (IGF2) was identified to be a direct target of miR-491-5p in CRC cells, and overexpression of IGF2 rescued the miR-491-5p-induced suppression of proliferation in CRC cells. Finally, we demonstrated that plasma miR-491-5p expression was decreased in CRC when compared to healthy controls and might be an effective diagnostic biomarker for CRC.
These data showed that miR-491-5p functioned as a tumor suppressor by targeting IGF2 in CRC, and miR-491-5p could serve as a potential diagnostic and prognostic biomarker for CRC.
微小RNA(miRNA)的失调在肿瘤发生中至关重要,据报道miR-491-5p的异常表达在各种癌症的发生和发展中起关键作用。然而,miR-491-5p在结直肠癌(CRC)中的生物学功能和潜在机制仍不清楚。
采用定量实时聚合酶链反应(qRT-PCR)评估CRC组织、细胞系和血浆中miR-491-5p和胰岛素样生长因子2(IGF2)mRNA的表达水平。使用细胞计数试剂盒-8和集落形成试验检测miR-491-5p对CRC细胞生长的影响。应用荧光素酶报告基因试验来确认miR-491-5p的靶基因。在裸鼠中进行体内实验。
发现miR-491-5p在CRC组织和细胞系中明显下调,且miR-491-5p表达水平降低与分化、TNM分期及总体生存率差相关。miR-491-5p过表达在体外和体内均抑制CRC细胞增殖。机制上,胰岛素样生长因子2(IGF2)被确定为CRC细胞中miR-491-5p的直接靶标,IGF2过表达挽救了miR-491-5p诱导的CRC细胞增殖抑制。最后,我们证明与健康对照相比,CRC患者血浆中miR-491-5p表达降低,且其可能是CRC的一种有效的诊断生物标志物。
这些数据表明,miR-491-5p在CRC中通过靶向IGF2发挥肿瘤抑制作用,且miR-491-5p可作为CRC潜在的诊断和预后生物标志物。