PO.CL12.03 · 临床研究
DNMT-1抑制剂的表观遗传调控诱导SK-N-AS神经母细胞瘤细胞死亡
Epigenetic modulation by DNMT-1 inhibitor induces cell death in SK-N-AS neuroblastoma cells
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
神经母细胞瘤（NB）是最常见的颅外实体瘤之一，通常见于新生儿和婴儿。不幸的是，大多数儿童在诊断时已表现为晚期疾病并有不良预后。尽管近期的治疗策略已显著改善了神经母细胞瘤的治疗，许多患者仍未从中获益。因此，迫切需要能够控制侵袭性和耐药性肿瘤的新治疗方法。通过DNA甲基化和组蛋白修饰进行的表观遗传调控已被证明在神经母细胞瘤进展中发挥关键作用，类似于在许多其他癌症类型中观察到的现象。在这方面，DNMTs在侵袭性神经母细胞瘤细胞中普遍上调，促进了肿瘤抑制通路的转录抑制。因此，在本研究中，我们探讨了DNMT/G9a抑制剂CM-272对SK-N-AS神经母细胞瘤细胞存活的影响。用2 µM浓度的CM-272处理神经母细胞瘤细胞导致显著的细胞死亡，这是由于凋亡标志物水平升高引起的程序性细胞死亡（PCD）级联的激活。此外，CM-272显著抑制DNMT-1表达，证实了其破坏DNA甲基化依赖性基因沉默的能力。在多种癌症类型中，凋亡诱导通过PARP裂解增强以及BAX和Caspases升高而得到验证。此外，CM-272处理升高了BAX表达，同时下调BCL-2水平，激活了内在通路。除上述分析外，还使用NanoString人miRNA面板进行了miRNA表达谱分析，以充分探索可能与CM-272处理相关的转录后调控机制。总之，这些结果表明CM-272能够通过靶向表观遗传调节因子和激活细胞死亡通路诱导SK-N-AS神经母细胞瘤细胞凋亡。我们的发现提示CM-272作为一种表观遗传修饰药物具有强大的治疗潜力，尤其是在对常规治疗耐药的神经母细胞瘤肿瘤中。致谢：本项目得到了国家儿科癌症基金会（NPCF）的支持。本研究还部分由佛罗里达州卫生部授予A.R.（通过迈阿密大学药理学系，佛罗里达州科勒尔盖布尔斯，美国）的Bankhead Coley基础设施发展基金资助。作者感谢皇家癌症研究女士协会（Royal Dames of Cancer Research Inc.，佛罗里达州劳德代尔堡）在开展本研究中提供的资金支持。
查看英文原文 English abstract
Neuroblastoma (NB) is one of the most common extra-cranial solid tumors that is typically found in newborns and infants. Unfortunately, most children are present with advanced diseases and have poor prognosis at the time of diagnosis. Although the treatment of neuroblastoma has significantly improved with recent therapeutic strategies, many patients still do not benefit from them. As a result, new treatment approaches that can control aggressive and resistant tumors are urgently needed. Epigenetic regulation through DNA methylation and histone modifications have been shown to play a critical role in neuroblastoma progression, similar to the phenomenon seen in many other types of cancers. In this regard, DNMT's are commonly upregulated in aggressive neuroblastoma cells, contributing to transcriptional repression of tumor suppressor pathways. Therefore, in this study, we investigated the effect of the DNMT/G9a inhibitor CM-272 on cell survival in SK-N-AS neuroblastoma cells. Treatment of neuroblastoma cells with CM-272 at a concentration of 2 µM resulted in significant cell death, due to activation of the programmed cell death (PCD) cascade caused by elevated levels of apoptotic markers. In addition, CM-272 markedly suppressed DNMT-1 expression, confirming its ability to disrupt DNA methylation dependent gene silencing. In multiple cancer types, apoptosis induction was validated by enhanced cleavage of PARP accompanied by elevation of BAX and Caspases. Furthermore, CM-272 treatment elevated BAX expression, with downregulated BCL-2 levels, Intrinsic Pathway activation. In addition to the above analyses, miRNA expression profiling was performed using the NanoString Human miRNA panel. To fully explore post-transcriptional regulatory mechanisms that may be associated with CM-272 treatment. Together, these results demonstrate that CM-272 can induces apoptosis in SK-N-AS neuroblastoma cells by targeting epigenetic regulators and activating cell death pathways. Our findings suggest that CM-272 has strong therapeutic potential, as an epigenetic modifier drug, particularly in neuroblastoma tumors that are resistant to conventional treatments. Acknowledgements: This project was supported by the National Pediatric Cancer Foundation (NPCF). This research was also partially funded by the Bankhead Coley Infrastructure Development Grant from the Florida Department of Health that was awarded to A.R. through the Department of Pharmacology of the University of Miami (Coral Gables, Florida USA). The authors would like to thank Royal Dames of Cancer Research Inc. (Ft. Lauderdale, Florida) for their financial support in conducting this research
利益披露 Disclosure
S. Jaganathan, None..
U. Natarajan, None..
A. Rathinavelu, None.