A genome-wide siRNA screen for regulators of tumor suppressor p53 activity in human non-small cell lung cancer cells identifies components of the RNA splicing machinery as targets for anticancer treatment.
A genome-wide siRNA screen for regulators of tumor suppressor p53 activity in human non-small cell lung cancer cells identifies components of the RNA splicing machinery as targets for anticancer treatment.
复制标题
针对人类非小细胞肺癌细胞中肿瘤抑制因子 p53 活性调节因子的全基因组 siRNA 筛选，确定了 RNA 剪接机制的组成部分作为抗癌治疗的靶标。
DOI：
10.1002/1878-0261.12052
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发表时间：
2017-05
影响因子：
6.6
通讯作者：
van Beusechem VW
中科院分区：
文献类型：
--
作者：
Siebring-van Olst E;Blijlevens M;de Menezes RX;van der Meulen-Muileman IH;Smit EF;van Beusechem VW
恢复野生型肿瘤抑制因子p53活性可能是治疗癌症的一个有价值的选择。为了促进非小细胞肺癌（NSCLC）新治疗方案的开发，我们对NSCLC细胞中p53活性的决定因素进行了全基因组siRNA筛选。我们发现了许多以前不知道参与调节p53活性的基因。沉默p53通路抑制剂基因与细胞活力的丧失相关。影响p53活性的最大功能基因簇是mRNA剪接。在特定剪接体成分沉默后观察到显着的p53激活，而不是通过剪接体的普遍抑制。在多种NSCLC细胞系中验证了10个基因作为p53活性的抑制剂：编码Ras通路激活剂SOS1、锌指蛋白TSHZ3、线粒体膜蛋白COX16和剪接体组分SNRPD3、SF3A3、SF3B1、SF3B6、XAB 2、CWC22和HNRNPL的基因。沉默这些基因通常会增加p53水平，对CDKN1A表达，诱导细胞周期停滞和细胞死亡有明显影响。沉默剪接体成分与MDM4 mRNA的选择性剪接相关，这可能有助于p53的激活。此外，沉默剪接因子在杀死NSCLC细胞方面特别有效，尽管是以p53非依赖性方式。有趣的是，沉默SNRPD3和SF3A3对NSCLC细胞比对肺成纤维细胞产生更强的细胞毒性，这表明这些基因可能代表有用的治疗靶点。
Reinstating wild‐type tumor suppressor p53 activity could be a valuable option for the treatment of cancer. To contribute to development of new treatment options for non‐small cell lung cancer (NSCLC), we performed genome‐wide siRNA screens for determinants of p53 activity in NSCLC cells. We identified many genes not previously known to be involved in regulating p53 activity. Silencing p53 pathway inhibitor genes was associated with loss of cell viability. The largest functional gene cluster influencing p53 activity was mRNA splicing. Prominent p53 activation was observed upon silencing of specific spliceosome components, rather than by general inhibition of the spliceosome. Ten genes were validated as inhibitors of p53 activity in multiple NSCLC cell lines: genes encoding the Ras pathway activator SOS1, the zinc finger protein TSHZ3, the mitochondrial membrane protein COX16, and the spliceosome components SNRPD3, SF3A3, SF3B1, SF3B6, XAB2, CWC22, and HNRNPL. Silencing these genes generally increased p53 levels, with distinct effects on CDKN1A expression, induction of cell cycle arrest and cell death. Silencing spliceosome components was associated with alternative splicing of MDM4 mRNA, which could contribute to activation of p53. In addition, silencing splice factors was particularly effective in killing NSCLC cells, albeit in a p53‐independent manner. Interestingly, silencing SNRPD3 and SF3A3 exerted much stronger cytotoxicity to NSCLC cells than to lung fibroblasts, suggesting that these genes could represent useful therapeutic targets.
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影响因子：
12.3
作者：
Boutros M;Brás LP;Huber W
通讯作者：
Huber W
影响因子：
3.2
作者：
Hong, Bo;van den Heuvel, A. Pieter J.;El-Deiry, Wafik S.
通讯作者：
El-Deiry, Wafik S.
影响因子：
8
作者：
Allende-Vega, N.;Dayal, S.;Saville, M. K.
通讯作者：
Saville, M. K.
影响因子：
21.3
作者：
Krastev, Dragomir B.;Slabicki, Mikolaj;Buchholz, Frank
通讯作者：
Buchholz, Frank
影响因子：
3.7
作者：
Laín, S;Midgley, C;Lane, DP
通讯作者：
Lane, DP