Flotillin-2 regulates epidermal growth factor receptor activation, degradation by Cbl-mediated ubiquitination, and cancer growth.
Flotillin-2 regulates epidermal growth factor receptor activation, degradation by Cbl-mediated ubiquitination, and cancer growth.
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Flotillin-2调节表皮生长因子受体的激活，CBL介导的泛素化降解和癌症生长。
DOI：
10.1016/j.jbc.2022.102766
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发表时间：
2023-01
影响因子：
4.8
通讯作者：
Lipkowitz, Stanley
中科院分区：
文献类型：
--
作者：
Wisniewski, David J.;Liyasova, Mariya S.;Korrapati, Soumya;Zhang, Xu;Ratnayake, Shashikala;Chen, Qingrong;Gilbert, Samuel F.;Catalano, Alexis;Voeller, Donna;Meerzaman, Daoud;Guha, Udayan;Porat-Shliom, Natalie;Annunziata, Christina M.;Lipkowitz, Stanley
表皮生长因子受体(EGFR)信号在多种癌症中经常发生异常。泛素连接酶Casitas B系淋巴瘤原癌基因(Cbl)通过泛素化调节活化的EGFR的降解，并作为适配器招募运输所需的蛋白质。在这里，我们使用细胞培养中氨基酸的稳定同位素标记质谱学来比较有或没有表皮生长因子(EGF)刺激的Cbl复合体。我们鉴定了100多个新的Cbl相互作用因子，二次siRNA筛选发现，Flotillin-2(FLOT2)的敲除导致EGF刺激下HeLa细胞中EGFR的磷酸化和降解增加。在PC9和H441细胞中，FLOT2基因敲除增加了EGF刺激的EGFR磷酸化、泛素化和下游信号转导，这可被EGFR抑制剂erlotinib逆转。在HeLa细胞中，FLOT2的CRISPR基因敲除(KO)证实了EGFR下调，信号增加，二聚化和内体转运增加。此外，我们确定FLOT2与Cb1和EGFR都有相互作用。EGFR对FLOT2丢失的下调是Cb1依赖的，因为Cb1和Cbl-b的协同敲除恢复了EGFR水平。此外，FLOT2过表达降低了EGFR信号转导和生长。在HEK293T细胞中，野生型(WT)FLOT2的过表达，而不是可溶性的G2AFLOT2突变体，抑制了EGF刺激下的EGFR磷酸化。FLOT2缺失可诱导EGFR依赖性增殖和锚定非依赖性生长。最后，FLOT2 KO可分别增加裸鼠和NSG小鼠的肿瘤形成率和肿瘤体积。综上所述，这些数据表明，FLOT2负面调控EGFR的激活和二聚化，以及随后的泛素化、内体运输和降解，导致体外和体内的增殖减少。
Epidermal growth factor receptor (EGFR) signaling is frequently dysregulated in various cancers. The ubiquitin ligase Casitas B-lineage lymphoma proto-oncogene (Cbl) regulates degradation of activated EGFR through ubiquitination and acts as an adaptor to recruit proteins required for trafficking. Here, we used stable isotope labeling with amino acids in cell culture mass spectrometry to compare Cbl complexes with or without epidermal growth factor (EGF) stimulation. We identified over a hundred novel Cbl interactors, and a secondary siRNA screen found that knockdown of Flotillin-2 (FLOT2) led to increased phosphorylation and degradation of EGFR upon EGF stimulation in HeLa cells. In PC9 and H441 cells, FLOT2 knockdown increased EGF-stimulated EGFR phosphorylation, ubiquitination, and downstream signaling, reversible by EGFR inhibitor erlotinib. CRISPR knockout (KO) of FLOT2 in HeLa cells confirmed EGFR downregulation, increased signaling, and increased dimerization and endosomal trafficking. Furthermore, we determined that FLOT2 interacted with both Cbl and EGFR. EGFR downregulation upon FLOT2 loss was Cbl dependent, as coknockdown of Cbl and Cbl-b restored EGFR levels. In addition, FLOT2 overexpression decreased EGFR signaling and growth. Overexpression of wildtype (WT) FLOT2, but not the soluble G2A FLOT2 mutant, inhibited EGFR phosphorylation upon EGF stimulation in HEK293T cells. FLOT2 loss induced EGFR-dependent proliferation and anchorage-independent growth. Lastly, FLOT2 KO increased tumor formation and tumor volume in nude mice and NSG mice, respectively. Together, these data demonstrated that FLOT2 negatively regulated EGFR activation and dimerization, as well as its subsequent ubiquitination, endosomal trafficking, and degradation, leading to reduced proliferation in vitro and in vivo.
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实现癌症基因组数据的共同愿景。
DOI：
10.1056/nejmp1607591
发表时间：
2016-09-22
期刊：
The New England journal of medicine
影响因子：
--
作者：
Grossman RL;Heath AP;Ferretti V;Varmus HE;Lowy DR;Kibbe WA;Staudt LM
通讯作者：
Staudt LM
影响因子：
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Lipkowitz S
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Dai, Zhijun
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作者：
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Tian, Ruijun
DOI：
10.1016/j.bbamcr.2014.04.013
发表时间：
2014-09-01
影响因子：
5.1
作者：
Asp, Nagham;Pust, Sascha;Sandvig, Kirsten
通讯作者：
Sandvig, Kirsten