Cancer Institute, Fudan University Shanghai Cancer Center, Shanghai, China.
Cancer Institute, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Cell Prolif. 2019 Mar;52(2):e12536. doi: 10.1111/cpr.12536. Epub 2018 Oct 19.
The present study aimed to reveal expression status of the neddylation enzymes in HNSCC and to elucidate the anticancer efficacy and the underlying mechanisms of inhibiting neddylation pathway.
The expression levels of neddylation enzymes were estimated by Western blotting in human HNSCC specimens and bioinformatics analysis of the cancer genome atlas (TCGA) database. Cell apoptosis was evaluated by Annexin V fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) stain and fluorescence-activated cell sorting (FACS). Small interfering RNA (siRNA) and the CRISPR-Cas9 system were used to elucidate the underlying molecular mechanism of MLN4924-induced HNSCC apoptosis.
Expression levels of NAE1 and UBC12 were prominently higher in HNSCC tissues than that in normal tissues. Inactivation of the neddylation pathway significantly inhibited malignant phenotypes of HNSCC cells. Mechanistic studies revealed that MLN4924 induced the accumulation of CRL ligase substrate c-Myc that transcriptionally activated pro-apoptotic protein Noxa, which triggered apoptosis in HNSCC.
These findings determined the over-expression levels of neddylation enzymes in HNSCC and revealed novel mechanisms underlying neddylation inhibition induced growth suppression in HNSCC cells, which provided preclinical evidence for further clinical evaluation of neddylation inhibitors (eg, MLN4924) for the treatment of HNSCC.
本研究旨在揭示 neddylation 酶在头颈部鳞状细胞癌（HNSCC）中的表达状态，并阐明抑制 neddylation 通路的抗癌疗效和潜在机制。
通过 Western blot 法检测人 HNSCC 标本和癌症基因组图谱（TCGA）数据库的生物信息学分析来评估 neddylation 酶的表达水平。通过 Annexin V 荧光素异硫氰酸酯/碘化丙啶（Annexin V-FITC/PI）染色和荧光激活细胞分选（FACS）评估细胞凋亡。使用小干扰 RNA（siRNA）和 CRISPR-Cas9 系统来阐明 MLN4924 诱导 HNSCC 细胞凋亡的潜在分子机制。
与正常组织相比，NAE1 和 UBC12 的表达水平在 HNSCC 组织中明显升高。neddylation 通路的失活显著抑制了 HNSCC 细胞的恶性表型。机制研究表明，MLN4924 诱导 CRL 连接酶底物 c-Myc 的积累，从而转录激活促凋亡蛋白 Noxa，引发 HNSCC 细胞凋亡。
这些发现确定了 neddylation 酶在 HNSCC 中的高表达水平，并揭示了 neddylation 抑制诱导 HNSCC 细胞生长抑制的新机制，为进一步临床评估 neddylation 抑制剂（如 MLN4924）治疗 HNSCC 提供了临床前证据。
Zotero 插件
