EGF/EGFR upregulates and cooperates with Netrin-4 to protect glioblastoma cells from DNA damage-induced senescence.
EGF/EGFR upregulates and cooperates with Netrin-4 to protect glioblastoma cells from DNA damage-induced senescence.
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EGF/EGFR上调并与Netrin-4配合保护胶质母细胞瘤细胞免受DNA损伤诱导的衰老
DOI：
10.1186/s12885-018-5056-4
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发表时间：
2018-12-04
期刊：
影响因子：
3.8
通讯作者：
Du Z
中科院分区：
文献类型：
--
作者：
Li L;Huang Y;Gao Y;Shi T;Xu Y;Li H;Hyytiäinen M;Keski-Oja J;Jiang Q;Hu Y;Du Z
研究背景多形性胶质母细胞瘤（GBM）是中枢神经系统恶性程度最高的肿瘤。烷化剂替莫唑胺（TMZ）是目前治疗GBM的一线化疗药物。然而，GBM细胞对TMZ的敏感性受到多种因素的影响。而且，一些临床试验，包括TMZ与其他药物的联合给药，都未能成功治疗GBM。我们以前曾报道，Netrin-4（NTN-4），层粘连蛋白样轴突导向蛋白，在TMZ引发的DNA损伤后GBM细胞衰老中起保护作用。然而，NTN 4的主调节器需要进一步阐明。表皮生长因子/表皮生长因子受体（Epidermal growth factor/Epidermal growth factor receptor，EGF/EGFR）可调节多种细胞外基质相关分子的表达，防止GBM细胞DNA损伤。在这项研究中，我们研究了EGF/EGFR信号和NTN 4之间的关系，并探讨其对TMZ treatment.MethodsCo-expression分析后，在GBM细胞的治疗效果进行使用NIH 934细胞系的RNA测序数据和从GBM肿瘤的单细胞RNA测序数据。共表达基因用于GO富集和信号传导途径富集。通过qPCR定量靶基因的mRNA表达，并通过衰老相关β-半乳糖苷酶染色研究细胞衰老。通过免疫印迹法观察和分析蛋白磷酸化。从TCGA-glioblastoma项目中提取TMZ治疗患者的RNA测序数据和临床信息，然后用于Kaplan-Meier生存分析。GO富集EGFR相关基因表明EGFR以类似于中枢神经系统发育和神经细胞分化的方式调节GBM细胞。通路分析表明EGFR及其相关基因参与细胞粘附、细胞外基质（ECM）组织化和caspase相关信号传导。我们还表明，EGF刺激GBM细胞中的NTN 4表达，并与NTN 4合作，可能通过AKT和ERK来减弱DNA损伤诱导的GBM细胞衰老。临床分析表明，EGFR和NTN 4的共同表达显著预测TMZ治疗GBM patients. Conclusions生存不良，本研究表明，EGF/EGFR调节并协同NTN 4在GBM中抵抗DNA损伤。因此，我们的发现为GBM提供了潜在的治疗靶点。
BackgroundGlioblastoma multiforme (GBM) is the most malignant central nervous system tumor. Alkylating agent, temozolomide (TMZ), is currently the first-line chemotherapeutic agent for GBM. However, the sensitivity of GBM cells to TMZ is affected by many factors. And, several clinic trials, including co-administration of TMZ with other drugs, have failed in successful treatment of GBM. We have previously reported that Netrin-4 (NTN4), a laminin-like axon guidance protein, plays a protective role in GBM cell senescence upon TMZ-triggered DNA damage. However, the master regulator of NTN4 needs further elucidation. Epidermal growth factor/Epidermal growth factor receptor (EGF/EGFR) can modulate the expression of various extracellular matrix related molecules, and prevent DNA damage in GBM cells. In this study, we investigated the relationship between EGF/EGFR signaling and NTN4, and explored their effect on therapeutic efficacy in GBM cells upon TMZ treatment.MethodsCo-expression analysis were performed by using the RNA sequencing data from NIH 934 cell lines and from single cell RNA sequencing data of GBM tumor. The co-expressing genes were used for GO enrichment and signaling pathway enrichment. mRNA expression of the target genes were quantified by qPCR, and cell senescence were investigated by Senescence-Associated Beta-Galactosidase Staining. Protein phosphorylation were observed and analyzed by immunoblotting. The RNA sequencing data and clinical information of TMZ treated patients were extracted from TCGA-glioblastoma project, and then used for Kaplan-Meier survival analysis.ResultsAnalysis of RNA sequencing data revealed a potential co-expression relationship betweenNTN4andEGFR. GO enrichment ofEGFR-correlated genes indicated that EGFR regulates GBM cells in a manner similar to that in central nervous system development and neural cell differentiation. Pathway analysis suggested thatEGFRand its related genes contribute to cell adhesion, extracellular matrix (ECM) organization and caspase related signaling. We also show that EGF stimulates NTN4 expression in GBM cells and cooperates with NTN4 to attenuate GBM cell senescence induced by DNA damage, possibly via AKT and ERK. Clinical analysis showed that co-expression of EGFR and NTN4 significantly predicts poor survival in TMZ-treated GBM patients.ConclusionsThis study indicates thatEGF/EGFRregulates and cooperates with NTN4 in DNA damage resistance in GBM. Therefore, our findings provide a potential therapeutic target for GBM.
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DOI：
10.1016/j.ijrobp.2012.10.008
发表时间：
2013-04-01
影响因子：
7
作者：
Chakravarti, Arnab;Wang, Meihua;Robins, H. Ian;Lautenschlaeger, Tim;Curran, Walter J.;Brachman, David G.;Schultz, Christopher J.;Choucair, Ali;Dolled-Filhart, Marisa;Christiansen, Jason;Gustavson, Mark;Molinaro, Annette;Mischel, Paul;Dicker, Adam P.;Bredel, Markus;Mehta, Minesh
通讯作者：
Mehta, Minesh
影响因子：
8.8
作者：
Günther, W;Pawlak, E;Damasceno, R;Arnold, H;Terzis, AJ
通讯作者：
Terzis, AJ
影响因子：
19
作者：
Klemm F;Joyce JA
通讯作者：
Joyce JA
外用 Netrin-4 抑制碱烧伤大鼠角膜新生血管的治疗作用
影响因子：
3.7
作者：
Han Y;Shao Y;Liu T;Qu YL;Li W;Liu Z
通讯作者：
Liu Z
影响因子：
158.5
作者：
Hegi, ME;Diserens, A;Stupp, R
通讯作者：
Stupp, R