NEDD8共价修饰抑制因子MLN4924对人卵巢癌细胞株SKOV3增殖和凋亡的影响  

Effects of NEDD8 covalent modification inhibitor MLN4924 on proliferation and apoptosis of human ovarian cancer cell line SKOV3

作　　者：刘健 吴会芳 赵亚伟 张纪岩(指导)[1] LIU Jian;WU Hui-Fang;ZHAO Ya-Wei;ZHANG Ji-Yan(Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学,南宁530021 [2]军事科学院,军事医学研究院军事认知与脑科学研究所,北京100850 [3]漯河市中心医院,漯河462000

出　　处：《中国免疫学杂志》2018年第3期340-343,348,共5页Chinese Journal of Immunology

基　　金：本文受北京市自然科学基金面上项目(5162026)、国家自然科学基金面上项目(81672368)和北京市自然科学基金(7174329)资助。

摘　　要：目的:探讨神经前体细胞发育相关受控表达分子8(NEDD8)共价修饰抑制因子MLN4924对人卵巢癌细胞增殖和凋亡的影响,并分析其可能机制。方法:使用不同浓度MLN4924(0、0.125、0.25、0.5μmol/L)处理人卵巢癌细胞株SKOV3 4 h,免疫印迹法检测PAR3、HER2、Neddylated-cullins、P-IκBα的表达水平;同时取细胞培养上清用ELISA检测IL-6的分泌。不同浓度MLN4924处理SKOV3细胞72 h,CCK8法检测细胞增殖抑制率,流式细胞仪检测细胞周期和细胞凋亡率。结果:MLN4924作用4 h时,能够使Neddylated-cullins下降、P-IκBα积聚,IL-6分泌减少,而PAR3、HER2表达水平变化不显著。MLN4924在0.125、0.25和0.5μmol/L的浓度下处理SKOV3细胞72 h时,对细胞增殖有明显的抑制作用,并且随着剂量的加大,抑制效果也显著提高;同时加药组S期细胞百分率相较于对照组明显升高,且形成大量四倍体,使之没有完整的细胞周期;细胞凋亡率也随MLN4924剂量增大而升高。结论:NEDD8修饰特异性抑制剂MLN4924能够明显抑制人卵巢癌细胞株SKOV3的增殖,诱导细胞周期阻滞和凋亡。上述效应可能与NF-κB通路活性受到抑制和IL-6表达下降有关。Objective:To explore the effects of neural precursor cell-expressed developmentally down regulated 8(NEDD8)covalent modification on ovarian cancer cell proliferation and apoptosis,and the possible underlying mechanisms.Methods:Use different concentrations of MLN4924(0,0.125,0.25,0.5 mol/L)and human ovarian cancer cell line SKOV3,the expression levels of PAR3,HER2,Neddylated-cullins,P-IκBαwere detected by Western blot and the secretion of IL-6 was measured by ELISA after 4 h treatment.The cell proliferation was determined by CCK-8 staining and the cell cycle and apoptosis were analyzed by flow cytometry after 72 h treatment.Results:MLN4924 inhibits the proliferation of SKOV3 cells in a dose-dependent manner,which is associated with reduced IL-6 secretion.Western blot revealed that MLN4924 dose-dependently inhibits the neddylation of cullins(reduced neddylated-cullins)and induced the accumulation of P-IκBα,whereas the expression levels of PARs and HER2 remained largely unchanged.At the same time MLN4924 dose-dependently induced cell cycle prove at S phase and the formation of a large number of tetraploids.Furthermore,apoptosis increased with the dose of MLN4924.Conclusion:NEDD8 covalent modification specific inhibitor MLN4924 can significantly inhibit the proliferation of SKOV3 cells and induce cell cycle arrest and apoptosis.The above inhibitory effects may partially result from impaired P-IκBαdegradation and IL-6 secretion.

关 键 词：MLN4924 卵巢癌细胞 细胞增殖 细胞凋亡 IL-6 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]