PA28gamma emerges as a novel functional target of tumour suppressor microRNA-7 in non-small-cell lung cancer.
PA28gamma emerges as a novel functional target of tumour suppressor microRNA-7 in non-small-cell lung cancer.
复制标题
PA28gamma 成为非小细胞肺癌中抑癌基因 microRNA-7 的新功能靶点。
DOI：
10.1038/bjc.2013.728
复制
发表时间：
2014-01-21
影响因子：
8.8
通讯作者：
Chu, Y.
中科院分区：
文献类型：
--
作者：
Xiong, S.;Zheng, Y.;Jiang, P.;Liu, R.;Liu, X.;Qian, J.;Gu, J.;Chang, L.;Ge, D.;Chu, Y.
MicroRNA-7（miR-7）是一种肿瘤抑制基因。然而，它是否在非小细胞肺癌（NSCLC）的生长中发挥作用以及它在肿瘤生长中的靶点仍在研究中。本研究采用非小细胞肺癌组织标本、非小细胞肺癌细胞系和组织芯片进行研究。总RNA、miRNA和蛋白质用于RT-PCR和western blot分析。免疫组化在组织芯片中进行。在体外和体内进行细胞培养和干预实验。通过生物信息学预测、western blot和荧光素酶检测，确定了miR-7的作用靶点。在本研究中，我们发现miR-7的表达不仅在NSCLC细胞系中显著下调，而且在人类NSCLC组织中与匹配的邻近组织相比也显著下调。在体外，通过miR-7模拟物在A549和H1299 NSCLC细胞中恢复其表达可抑制细胞增殖、集落形成和细胞周期进展。更重要的是，在体内施用miR-7后，裸鼠的致瘤性降低。在此基础上，通过生物信息学分析、荧光素酶分析和蛋白质印迹分析，我们确定了一个新的miR-7靶点PA 28 gamma（一种蛋白酶体激活剂）参与肿瘤的调控。PA 28 γ mRNA和蛋白水平在NSCLC细胞系和肿瘤样本中显著上调，与miR-7呈强负相关。此外，在NSCLC细胞中，PA 28 γ的敲低诱导了与miR-7过表达相似的效应。此外，miR-7过表达或PA 28 γ沉默可降低NSCLC细胞系中cyclinD 1在mRNA和蛋白水平的表达。所有这些发现都有力地表明，NSCLC中miR-7表达下调导致的PA 28 γ过表达在促进癌细胞进展中具有重要作用，从而导致NSCLC生长。因此，靶向PA 28 γ和/或miR-7的策略可能成为NSCLC治疗中有前途的分子疗法。
MicroRNA-7 (miR-7) has been reported to be a tumour suppressor gene. However, whether it has a role in the growth of non-small-cell lung cancer (NSCLC) and what is its target involved in the tumour growth is still under investigation. NSCLC tissue sample, NSCLC cell lines and tissue microarray were investigated in this study. Total RNA, miRNA and protein were used for RT-PCR and western blot analysis. Immunohistochemistry was performed in tissues microarray. Cell culture and intervention experiments were performed in vitro and in vivo. Bioinformatics prediction, western blot and luciferase assay were identified the target of miR-7. In this study, we found that the expression of miR-7 was significantly downregulated not only in NSCLC cell lines, but also in human NSCLC tissues compared with the matched adjacent tissues. Restoration of its expression through miR-7 mimics in A549 and H1299 NSCLC cells inhibited cell proliferation, colony formation, and cell-cycle progression in vitro. More importantly, the tumorigenicity in nude mice was reduced after administration of miR-7 in vivo. In advance, through bioinformatic analysis, luciferase assay and western blot, we identified a novel target of miR-7, PA28gamma (a proteasome activator) to be enrolled in the regulation with tumour. PA28gamma mRNA and protein levels are markedly upregulated in NSCLC cell lines and tumour samples, exhibiting a strong inverse relation with that of miR-7. In addition, knockdown of PA28gamma induced similar effects as overexpression of miR-7 in NSCLC cells. Furthermore, miR-7 overexpression or silencing of PA28gamma reduced the cyclinD1 expression at mRNA and protein level in NSCLC cell lines. All these findings strongly imply that the overexpression of PA28gamma resulted from miR-7 downexpression in NSCLC has an important role in promoting cancer cell progress and consequently results in NSCLC growth. Thus, strategies targeting PA28gamma and/or miR-7 may become promising molecular therapies in NSCLC treatment.
登录
查看更多内容
影响因子：
11.2
作者：
Bader AG;Brown D;Winkler M
通讯作者：
Winkler M
DOI：
10.1165/rcmb.2010-0323oc
发表时间：
2011-08-01
影响因子：
6.4
作者：
Cushing, Leah;Kuang, Ping Ping;Lue, Jining
通讯作者：
Lue, Jining
影响因子：
64.5
作者：
Lamb, J;Ramaswamy, S;Ewen, ME
通讯作者：
Ewen, ME
影响因子：
4.4
作者：
Barton, LF;Runnels, HA;Monaco, JJ
通讯作者：
Monaco, JJ
影响因子：
20.4
作者：
Duncavage, Eric;Goodgame, Boone;Pfeifer, John
通讯作者：
Pfeifer, John