The impact of miR-34a on protein output in hepatocellular carcinoma HepG2 cells
The impact of miR-34a on protein output in hepatocellular carcinoma HepG2 cells
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DOI：
10.1002/pmic.200900646
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发表时间：
2010-04-01
期刊：
影响因子：
3.4
通讯作者：
Zheng, Shu-Sen
中科院分区：
文献类型：
--
作者：
Cheng, Jun;Zhou, Lin;Zheng, Shu-Sen
MicroRNA是一种小的非编码RNA分子，在从细胞周期到细胞迁移和侵袭的生物学过程中发挥重要作用。越来越多的证据表明，miR-34 a作为p53肿瘤抑制的关键介质，在人类癌症中异常表达。在本研究中，我们的目的是探索miR-34 a的精确生物学作用以及瞬时转染miR-34 a的肝癌细胞系HepG 2细胞中的整体蛋白变化。miR-34 a转染HepG 2细胞后，细胞增殖受到抑制，细胞迁移和侵袭能力受到抑制。它还诱导HepG 2细胞聚集在Cl期。在2-DE分离的116个差异表达蛋白点中，MALDI-TOF/TOF分析成功鉴定了34个蛋白。其中，15个下调蛋白可能是miR-34 a的下游靶点。生物信息学分析构建了一个蛋白质-蛋白质相互作用网络，发现p53信号通路和细胞周期通路是两个主要的枢纽，包含了大部分受miR-34 a调控的蛋白质。细胞骨架蛋白如LMNA、GFAP、MACF 1、ALDH 2和LOC 100129335是miR-34 a的潜在靶点。总之，miR-34 a功能的废除可能导致下游分子打开或关闭，导致HCC的发展。
MicroRNAs are small non-coding RNA molecules that play essential roles in biological processes ranging from cell cycle to cell migration and invasion. Accumulating evidence suggests that miR-34a, as a key mediator of p53 tumor suppression, is aberrantly expressed in human cancers. In the present study, we aimed to explore the precise biological role of miR-34a and the global protein changes in HCC cell line HepG2 cells transiently transfected with miR-34a. Transfection of miR-34a into HepG2 cells caused suppression of cell proliferation, inhibition of cell migration and invasion. It also induced an accumulation of HepG2 cells in Cl phase. Among 116 protein spots with differential expression separated by 2-DE method, 34 proteins were successfully identified by MALDI-TOF/TOF analysis. Of these, 15 down-regulated proteins may be downstream targets of miR-34a. Bioinformatics analysis produced a protein-protein interaction network, which revealed that the p53 signaling pathway and cell cycle pathway were two major hubs containing most of the proteins regulated by miR-34a. Cytoskeletal proteins such as LMNA, GFAP, MACF1, ALDH2, and LOC100129335 are potential targets of miR-34a. In conclusion, abrogation of miR-34a function could cause downstream molecules to switch on or off, leading to HCC development.