NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase.
NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase.
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DOI：
10.1186/s13045-017-0392-4
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发表时间：
2017-01-13
影响因子：
28.5
通讯作者：
Zhan F
中科院分区：
文献类型：
--
作者：
Gu Z;Xia J;Xu H;Frech I;Tricot G;Zhan F
有氧糖酵解是癌症的一个标志，其特征是在正常情况下葡萄糖代谢和乳酸生成增加。近年来，丙酮酸激酶M2 （PKM2）被认为是调节有氧糖酵解和促进肿瘤细胞增殖和存活的关键分子。采用串联亲和纯化、质谱联用（TAP-MS）和共免疫沉淀（Co-IP）技术研究了NIMA （never in mitosis gene A）相关激酶2 （NEK2）与异质核核糖核蛋白（hnRNP） A1/2之间的相互作用。采用RNA免疫沉淀法（RIP）鉴定NEK2与PKM前mrna序列的结合。采用染色质免疫沉淀(ChIP)-PCR分析c-Myc对NEK2的转录调控。Western blot和real-time PCR分析NEK2对PKM2的调控作用。NEK2通过与hnRNPA1/2相互作用调节多发性骨髓瘤细胞中PKM未成熟RNA的选择性剪接。RIP显示NEK2结合PKM pre-mRNA第9外显子侧翼的内含子序列。NEK2的敲低会降低PKM2/PKM1以及其他有氧糖酵解基因（包括GLUT4、HK2、ENO1、LDHA和MCT4）的比例。NEK2和PKM2高表达的骨髓瘤患者无事件生存期和总生存期较低。我们的数据表明NEK2在骨髓瘤细胞中受c-Myc的转录调节。NEK2的异位表达部分缓解了由沉默的c-Myc诱导的生长抑制和细胞死亡。我们的研究表明NEK2通过调节PKM剪接和增加骨髓瘤细胞中PKM2/PKM1的比例来促进有氧糖酵解，这有助于其致癌活性。
Aerobic glycolysis, a hallmark of cancer, is characterized by increased metabolism of glucose and production of lactate in normaxia. Recently, pyruvate kinase M2 (PKM2) has been identified as a key player for regulating aerobic glycolysis and promoting tumor cell proliferation and survival. Tandem affinity purification followed up by mass spectrometry (TAP-MS) and co-immunoprecipitation (Co-IP) were used to study the interaction between NIMA (never in mitosis gene A)-related kinase 2 (NEK2) and heterogeneous nuclear ribonucleoproteins (hnRNP) A1/2. RNA immunoprecipitation (RIP) was performed to identify NEK2 binding to PKM pre-mRNA sequence. Chromatin-immunoprecipitation (ChIP)-PCR was performed to analyze a transcriptional regulation of NEK2 by c-Myc. Western blot and real-time PCR were executed to analyze the regulation of PKM2 by NEK2. NEK2 regulates the alternative splicing of PKM immature RNA in multiple myeloma cells by interacting with hnRNPA1/2. RIP shows that NEK2 binds to the intronic sequence flanking exon 9 of PKM pre-mRNA. Knockdown of NEK2 decreases the ratio of PKM2/PKM1 and also other aerobic glycolysis genes including GLUT4, HK2, ENO1, LDHA, and MCT4. Myeloma patients with high expression of NEK2 and PKM2 have lower event-free survival and overall survival. Our data indicate that NEK2 is transcriptionally regulated by c-Myc in myeloma cells. Ectopic expression of NEK2 partially rescues growth inhibition and cell death induced by silenced c-Myc. Our studies demonstrate that NEK2 promotes aerobic glycolysis through regulating splicing of PKM and increasing the PKM2/PKM1 ratio in myeloma cells which contributes to its oncogenic activity.