PO.CL12.03 · 临床研究
MDM2和DNMT抑制剂介导的表观遗传调控促进神经母细胞瘤细胞的细胞周期阻滞与程序性细胞死亡
Epigenetic modulation by MDM2 and DNMT inhibitor promotes cell cycle arrest and programmed cell death in neuroblastoma cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
神经母细胞瘤是一种主要影响5岁以下儿童的儿科恶性肿瘤。该恶性肿瘤起源于神经嵴细胞（称为神经母细胞），可见于肾上腺、颈部、胸部或盆腔区域。尽管有多种治疗方法可用于治疗神经母细胞瘤，实验及文献证据提示表观遗传修饰可触发细胞周期阻滞并诱导程序性细胞死亡（PCD），如凋亡，从而为在神经母细胞瘤治疗中取得更大成功提供了潜在的治疗途径。我们的研究探讨了MDM2和DNMT抑制剂是否能够阻断DNMTs，并同时上调TET和p21，以在IMR-32和SK-N-AS细胞中诱导细胞周期阻滞和凋亡性细胞死亡。为评估RG-7388、CM-272和SGI-1027的作用，将神经母细胞瘤细胞处理24小时，随后进行qRT-PCR分析以评估DNMT-1、DNMT-3A、DNMT-3B、G9a、EZH2、TET-2、TET-3、MDM2、p53、p21和PARP的基因表达水平。此外，通过western blot方法评估上述基因的蛋白表达水平，以量化表观遗传、细胞周期及凋亡相关生物标志物的变化。有趣的是，qRT-PCR分析显示，与对照相比，CM-272和SGI-1027处理的SK-N-AS细胞中DNMT-1、DNMT-3A、DNMT-3B、G9a和EZH2水平下调。在IMR-32和SK-N-AS细胞中，western blot分析均显示药物处理后DNMT-1下调、PARP裂解以及BAX上调。迄今为止，我们的发现表明RG-7388、CM-272和SGI-1027处理有效下调了IMR-32细胞中DNMTs的表达，同时上调p21水平，导致细胞周期阻滞和凋亡。这些实验性化合物诱导细胞周期阻滞和细胞死亡的能力提示，对这些化合物的进一步评估可能有助于发现治疗侵袭性神经母细胞瘤的有前景的治疗药物。（本项目由国家儿科癌症基金会（NPCF）以及佛罗里达州劳德代尔堡的皇家癌症研究女士会（The Royal Dames of Cancer Research Inc.）资助）
查看英文原文 English abstract
Neuroblastoma isa pediatricmalignancythat affects childrenmostly under the age of 5 yearsold. Thismalignancyoriginatesfrom neural crest cells (called neuroblasts) that can be found inthe adrenal glands, neck, chest, or pelvicareas. Although varioustreatments are availablefortreatingneuroblastoma, experimentaland literatureevidencesuggest that epigeneticmodificationscan triggercell cycle arrest and induceprogrammed cell death (PCD), such asapoptosis,providinga potentialtherapeuticavenue for achievinggreater success duringneuroblastoma treatments. Our study investigatedwhether the MDM2 and DNMT inhibitorscould block DNMTs, and concurrently upregulate TET and p21, to inducecell cycle arrest andapoptoticcell death inIMR-32 and SK-N-AS cells. To assess the effects of RG-7388, CM-272,and SGI-1027, the neuroblastoma cells were treated for 24 hrs, then qRT-PCR analyses wereconducted to evaluate the gene expressionlevels of DNMT-1, DNMT-3A, DNMT-3B, G9a,EZH2, TET-2, TET-3, MDM2, p53, p21, and PARP. In addition,the proteinexpressionlevels ofthe previouslylistedgenes were evaluated through western blot methods to quantifythe changesinepigenetic,cell cycle, and apoptosis-relatedbiomarkers.Interestingly, the qRT-PCR analysisshowed a downregulationof DNMT-1, DNMT-3A, DNMT-3B, G9a, and EZH2 levels intheCM-272 and SGI-1027 treated SK-N-AS cells compared to the control. In both IMR-32 andSK-N-AS cells, the western blot analysisrevealed DNMT-1 downregulation,cleavage of PARP,and upregulationof BAX followingthe drug treatments. So far, our findingsindicatethatRG-7388, CM-272, and SGI-1027 treatments effectivelydownregulate the expressionof
DNMTs, whileupregulatingp21 levels inIMR-32 cells, leadingto cell cycle arrest andapoptosis.The abilityof these experimentalcompounds to inducecell cycle arrest and cell deathsuggests that further evaluationof these compounds could lead to the identificationof apromisingtherapeuticagent for treatingaggressiveneuroblastoma.(Thisprojectwas supported by the NationalPediatricCancer Foundation(NPCF) and The RoyalDames of Cancer Research Inc., Ft. Lauderdale, Florida)
利益披露 Disclosure
S. Talanki, None..
H. Le, None..
A. Chhabra, None..
R. Hede, None..
J. Nazario, None..
P. Srivastava, None..
G. Davu, None..
S. Jaganathan, None..
U. Natarajan, None..
A. Rathinavelu, None.