Lian Hai-Wei, Zhou Yun, Jian Zhi-Hong, Liu Ren-Zhong
Department of Neurosurgery, Wuhan University Renmin Hospital, Wuhan, China E-mail :
Asian Pac J Cancer Prev. 2014;15(23):10181-5. doi: 10.7314/apjcp.2014.15.23.10181.
MicroRNAs, small noncoding RNA molecules, can regulate mammalian cell growth, apoptosis and differentiation by controlling the expression of target genes. The aim of this study was to investigate the function of miR-323-5p in the glioma cell line, U251.
After over-expression of miR-323- 5p using miR-323-5p mimics, cell growth, apoptosis and migration were tested by MTT, flow cytometry and cell wound healing assay, respectively. We also assessed the influence of miR-323-5p on the mRNA expression of IGF- 1R by quantitative real-time reverse transcriptase PCR (qRT-PCR), and on the protein levels by Western blot analysi. In addition, dual-luciferase reporter assays were performed to determine the target site of miR-323-5p to IGF-1R 3'UTR.
Our findings showed that over-expression of miR-323-5p could promote apoptosis of U251 and inhibit the proliferation and migration of the glioma cells.
This study demonstrated that increased expression of miR-323-5p might be related to glioma progression, which indicates a potential role of miR-323-5p for clinical therapy.
微小RNA是一类小的非编码RNA分子,可通过控制靶基因的表达来调节哺乳动物细胞的生长、凋亡和分化。本研究旨在探讨miR-323-5p在胶质瘤细胞系U251中的功能。
使用miR-323-5p模拟物过表达miR-323-5p后,分别通过MTT法、流式细胞术和细胞划痕愈合试验检测细胞生长、凋亡和迁移情况。我们还通过定量实时逆转录聚合酶链反应(qRT-PCR)评估miR-323-5p对IGF-1R mRNA表达的影响,并通过蛋白质印迹分析评估其对蛋白质水平的影响。此外,进行双荧光素酶报告基因测定以确定miR-323-5p对IGF-1R 3'UTR的靶位点。
我们的研究结果表明,miR-323-5p的过表达可促进U251细胞凋亡,并抑制胶质瘤细胞的增殖和迁移。
本研究表明,miR-323-5p表达增加可能与胶质瘤进展有关,这表明miR-323-5p在临床治疗中具有潜在作用。