核因子-κB抑制剂联合核因子E2相关因子2抑制剂对人结肠癌HCT116细胞凋亡的作用机制研究  

Effect and mechanism of nuclear factor-κB inhibitor combined with nuclear factor E2 related factor 2 inhibitor on apoptosis of human colon cancer HCT116 cells

作　　者：罗欢 邹攀 魏行云 刘灵 何迎春[2,3] 江志超[3,4] LUO Huan;ZOU Pan;WEI Xing-yun;LIU Ling;HE Ying-chun;JIANG Zhi-chao(Post-Graduate School,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China;Hunan Provincial Key Laboratory for The Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Traditional Chinese Medicine and Visual Function Protection Engineering and Technological Research Center,Hunan University of Chinese Medicine,Changsha 410208,Hunan Province,China;The Second People’s Hospital of Hunan Province(Hunan Provincial Brain Hospital),Changsha 410007,Hunan Province,China)

机构地区：[1]湖南中医药大学研究生院,湖南长沙410208 [2]湖南中医药大学中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心,湖南长沙410208 [4]湖南省第二人民医院(湖南省脑科医院),湖南长沙410007

出　　处：《中国临床药理学杂志》2022年第2期123-126,141,共5页The Chinese Journal of Clinical Pharmacology

基　　金：湖南省自然科学基金资助项目(2018JJ2219)。

摘　　要：目的探究核因子κB(NF-κB)抑制剂(BAY11-7082)联合核因子E2相关因子2(Nrf2)抑制剂(ML385)对人结肠癌细胞HCT116的作用机制。方法体外培养人结肠癌细胞HCT116,将细胞分为4组:空白对照组、NF-κB抑制剂组、Nrf2抑制剂和联合组。空白对照组以RPMI-1640培养基进行培养,NF-κB抑制剂组以10μmol·L^(-1)BAY 11-7082干预,Nrf2抑制剂组以5μmol·L^(-1)ML385干预,联合组以10μmol·L^(-1)BAY 11-7082+5μmol·L^(-1)ML385干预,干预时间为24 h。MTT法检测人结肠癌细胞HCT116增殖抑制率,以流式细胞术和Hoechst 33342染色法检测细胞凋亡状况,以蛋白质印迹法检测凋亡相关蛋白及Nrf2、NF-κB通路相关蛋白表达水平。结果ML385、BAY 11-7082均能显著抑制人结肠癌细胞HCT116增殖(均P<0.05);干预24 h,空白对照组、Nrf2抑制剂组、NF-κB抑制剂组和联合组的细胞凋亡率分别为(1.77±0.60)%,(19.41±1.87)%,(21.55±2.33)%和(39.84±1.22)%;这4组的B细胞淋巴瘤-2相关X蛋白(Bax)的相对表达量分别为1.00,1.53±0.28,1.43±0.30和3.47±0.70;这4组的生存素蛋白的相对表达量分别为1.00,0.60±0.11,0.32±0.10和0.32±0.12;这4组的X连锁凋亡抑制蛋白(XIAP)的相对表达量分别为1.00,0.50±0.11,0.19±0.06和0.25±0.04。空白对照组、Nrf2抑制剂组和NF-κB抑制剂组的NF-κB蛋白的相对表达量分别为1.00,0.71±0.10和0.44±0.06;这3组的IκB蛋白分别为1.00,0.37±0.15和0.33±0.13;这3组的Nrf2蛋白分别为1.00,0.14±0.09和0.75±0.08。Nrf2抑制剂组、NF-κB抑制剂组和联合组与空白对照组比较,上述指标的差异均有统计学意义(均P<0.05)。结论NF-κB抑制剂联合Nrf2抑制剂可协同诱导人结肠癌细胞HCT116凋亡,该效应可能与下调Survivin和XIAP的表达、上调Bax表达有关。Objective To explore the effect of nuclear factor κB(NF-κB) inhibitor(BAY11-7082)combined with nuclear factor E2-related factor 2(Nrf2) inhibitor(ML385)on the apoptosis of human colon cancer HCT116 cells, and to explore the mechanism of action from the interaction of signal pathways. Methods Human colon cancer HCT116 cells were cultured in vitro. They were divided into 4 groups: Blank control group,NF-κB inhibitor group,Nrf2 inhibitor,and combined group. The blank control group was cultured in RPMI-1640 medium,and NF-κB inhibitor group was treated with 10 μmol·L^(-1) BAY 11-7082,Nrf2 inhibitor group was treated with 5 μmol·L^(-1) ML385,and combined group was treated with 10 μmol·L^(-1) BAY 11-7082 + 5μmol·L^(-1) ML385,for 24 h. MTT method was used to detect the proliferation inhibition rate of human colon cancer HCT116 cells. Hoechst 33342 staining method and flow cytometry were used to detect cell apoptosis. Western blot method was used to detect the protein expression levels of apoptosis-related proteins and Nrf2 and NF-κB pathway-related proteins. Results Both ML385 and BAY 11-7082 can inhibit the proliferation of human colon cancer HCT116 cells( all P < 0. 05). At 24 h of intervention,the apoptosis rates in blank control group,Nrf2 inhibitor group,NF-κB inhibitor group and combined group were( 1. 77 ± 0. 60) %,( 19. 41 ± 1. 87) %,( 21. 55 ± 2. 33) %,( 39. 84 ±1. 22) %;the relative expression levels of B-cell lymphoma-2 related X protein( Bax) in these four groups were 1. 00,1. 53 ± 0. 28,1. 43 ± 0. 30,3. 47 ± 0. 70;the relative expression of Survivin protein in these 4 groups were 1. 00,0. 60 ± 0. 11,0. 32 ± 0. 10,0. 32 ± 0. 12;the relative expression of X-linked inhibitor of apoptosis protein( XIAP) protein in these 4 groups were 1. 00,0. 50 ±0. 11,0. 19 ±0. 06,0. 25 ±0. 04. The relative expression of NF-κB protein in blank control group,Nrf2 inhibitor group and NF-κB inhibitor group were 1. 00,0. 71 ± 0. 10,0. 44 ± 0. 06;the relative expression levels of IκB protein in these

关 键 词：人结肠癌细胞 核因子-ΚB信号通路 核因子E2相关因子2信号通路 凋亡 协同 

分 类 号：R97[医药卫生—药品]