MicroRNA-34a is a potent tumor suppressor molecule in vivo in neuroblastoma.
MicroRNA-34a is a potent tumor suppressor molecule in vivo in neuroblastoma.
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DOI：
10.1186/1471-2407-11-33
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发表时间：
2011-01-25
期刊：
影响因子：
3.8
通讯作者：
Stallings RL
中科院分区：
文献类型：
--
作者：
Tivnan A;Tracey L;Buckley PG;Alcock LC;Davidoff AM;Stallings RL
神经母细胞瘤是一种源自交感神经系统前体细胞的儿科癌症，占儿童癌症死亡率的 15%。关于 miRNA 在神经母细胞瘤中的作用，miR-34a（映射到通常被删除的染色体 1p36 区域）被发现通过靶向与细胞增殖和凋亡相关的众多基因来充当肿瘤抑制因子。将合成的 miR-34a（或阴性对照）前体分子转染至 NB1691luc 和 SK-N-ASluc 神经母细胞瘤细胞中。在体外和体内分析之前，使用定量 PCR 来验证 NB1691luc 和 SK-N-ASluc 细胞系中 miR-34a 水平的增加。对 miR-34a 过度表达对细胞生长、细胞周期和信号转导途径中磷蛋白激活的影响进行了体外分析。将过度表达 miR-34a 的神经母细胞瘤细胞腹膜后注射到免疫受损的 CB17-SCID 小鼠中，并通过测量生物发光（光子/秒/cm2）来评估 21 天的肿瘤负荷。 72 小时内，NB1691luc 和 SK-N-ASluc 神经母细胞瘤细胞系中 miR-34a 的过度表达导致细胞数量相对于 premiR 阴性对照处理的细胞显着减少。流式细胞术结果表明，miR-34a 诱导细胞周期停滞和随后的细胞凋亡激活。磷蛋白分析强调了参与信号转导的关键元件，由于 miR-34a 引入细胞，信号转导的激活失调。作为 miR-34a 对磷蛋白水平作用的潜在机制，我们证明 miR-34a 过度表达会导致 MAP3K9 mRNA 和蛋白水平显着降低。尽管 MAP3K9 是 miR-34a 的预测靶标，但直接靶向无法通过荧光素酶报告基因检测进行验证。尽管如此，无论所涉及的机制如何，预计 miR-34a 导致的 MAP3K9 表达减少的任何功能效应都是相似的。最值得注意的是，体内研究表明，在腹膜后神经母细胞瘤中给予外源性 miR-34a 后，肿瘤生长受到显着抑制。我们首次证明 miR-34a 在神经母细胞瘤的体内原位小鼠模型中显着降低肿瘤生长，并确定了 miR-34a 对与细胞凋亡相关的关键蛋白的磷酸化激活的新作用。
Neuroblastoma is a paediatric cancer which originates from precursor cells of the sympathetic nervous system and accounts for 15% of childhood cancer mortalities. With regards to the role of miRNAs in neuroblastoma, miR-34a, mapping to a chromosome 1p36 region that is commonly deleted, has been found to act as a tumor suppressor through targeting of numerous genes associated with cell proliferation and apoptosis. A synthetic miR-34a (or negative control) precursor molecule was transfected into NB1691luc and SK-N-ASluc neuroblastoma cells. Quantitative PCR was used to verify increased miR-34a levels in NB1691luc and SK-N-ASluc cell lines prior to in vitro and in vivo analysis. In vitro analysis of the effects of miR-34a over expression on cell growth, cell cycle and phosphoprotein activation in signal transduction pathways was performed. Neuroblastoma cells over expressing miR-34a were injected retroperitoneally into immunocompromised CB17-SCID mice and tumor burden was assessed over a 21 day period by measuring bioluminescence (photons/sec/cm2). Over expression of miR-34a in both NB1691luc and SK-N-ASluc neuroblastoma cell lines led to a significant decrease in cell number relative to premiR-negative control treated cells over a 72 hour period. Flow cytometry results indicated that miR-34a induced cell cycle arrest and subsequent apoptosis activation. Phosphoprotein analysis highlighted key elements involved in signal transduction, whose activation was dysregulated as a result of miR-34a introduction into cells. As a potential mechanism of miR-34a action on phosphoprotein levels, we demonstrate that miR-34a over-expression results in a significant reduction of MAP3K9 mRNA and protein levels. Although MAP3K9 is a predicted target of miR-34a, direct targeting could not be validated with luciferase reporter assays. Despite this fact, any functional effects of reduced MAP3K9 expression as a result of miR-34a would be expected to be similar regardless of the mechanism involved. Most notably, in vivo studies showed that tumor growth was significantly repressed after exogenous miR-34a administration in retroperitoneal neuroblastoma tumors. We demonstrate for the first time that miR-34a significantly reduces tumor growth in an in vivo orthotopic murine model of neuroblastoma and identified novel effects that miR-34a has on phospho-activation of key proteins involved with apoptosis.
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影响因子：
--
作者：
Liu H;Brannon AR;Reddy AR;Alexe G;Seiler MW;Arreola A;Oza JH;Yao M;Juan D;Liou LS;Ganesan S;Levine AJ;Rathmell WK;Bhanot GV
通讯作者：
Bhanot GV
DOI：
10.1165/ajrcmb.26.5.4689
发表时间：
2002-05-01
影响因子：
6.4
作者：
Greenberg, AK;Basu, S;Lee, TC
通讯作者：
Lee, TC
microRNA miR-34抑制人类胰腺癌肿瘤发射细胞。
影响因子：
3.7
作者：
Ji Q;Hao X;Zhang M;Tang W;Yang M;Li L;Xiang D;Desano JT;Bommer GT;Fan D;Fearon ER;Lawrence TS;Xu L
通讯作者：
Xu L
影响因子：
4.7
作者：
Chim, C. S.;Wong, K. Y.;Liang, R.
通讯作者：
Liang, R.
影响因子：
9.7
作者：
Li, Na;Fu, Hanjiang;Zheng, Xiaofei
通讯作者：
Zheng, Xiaofei