土贝母苷乙加速NRF2蛋白降解促进结直肠癌细胞的凋亡  

Tubeimoside Ⅱ accelerates the degradation of NRF2 to promote apoptosis of colorectal cancer cell

作　　者：许微 张春云 李泽彦 伊娜娜 李枢媛 李雪[1] 常欣欣 张琳 高泽涛 杨清竹 XU Wei;ZHANG Chunyun;LI Zeyan;YI Nana;LI Shuyuan;LI Xue;CHANG Xinxin;ZHANG Lin;GAO Zetao;YANG Qingzhu(School of Life Sciences,Agriculture and Forestry,Qiqihar University,Qiqihar 161006,China;Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas,Qiqihar University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔大学,黑龙江省抗性基因工程与寒地多样性保护重点实验室,黑龙江齐齐哈尔161006

出　　处：《高师理科学刊》2024年第11期63-70,共8页Journal of Science of Teachers＇College and University

基　　金：2023年齐齐哈尔大学研究生创新科研项目(QUZLTS_CX2023010);黑龙江省大学生创新创业训练计划资助项目(S202410232014,X202410232120,x202310232036)。

摘　　要：探讨土贝母苷乙(Tubeimoside Ⅱ,TBMS Ⅱ)通过促进核转录因子红系2相关因子2(Transcription factor nuclear erythroid factor 2-like,NRF2)的降解进而影响结直肠癌细胞LOVO的死亡.培养人结直肠癌LOVO细胞,通过细胞活力实验检测TBMS Ⅱ对LOVO细胞的抑制作用,计算半致死率IC_(50)值,并根据IC_(50)值分组;克隆形成实验检测细胞克隆形成能力的影响;流式细胞仪检测细胞凋亡、细胞周期以及细胞中ROS水平的影响;生化试剂盒检测TBMS Ⅱ对细胞中还原型谷胱甘肽(Glutathione,GSH)、超氧化物歧化酶(Superoxide dismutase,SOD)、总抗氧化能力(Total antioxidant capacity,T-AOC)及丙二醛(Malondialdehyde,MDA)水平的影响;Western blot实验检测TBMS Ⅱ对LOVO细胞中NRF2蛋白表达的影响;分子对接探究TBMS Ⅱ和NRF2之间的结合能力.结果表明,TBMS Ⅱ显著抑制LOVO细胞的存活率和克隆形成能力,影响细胞周期,并诱导LOVO细胞的凋亡;TBMS Ⅱ抑制细胞中抗氧化指标GSH,SOD以及T-AOC的水平,上调细胞中MDA,ROS的水平;TBMS Ⅱ可能通过直接与NRF2结合,抑制细胞中NRF2蛋白的表达,促进细胞中NRF2蛋白的降解;抑制NRF2的表达能够增强TBMS Ⅱ诱导细胞凋亡的能力.因此,TBMS Ⅱ通过促进NRF2蛋白的降解诱导结直肠癌LOVO细胞的凋亡.To investigate the impact of Tubeimoside Ⅱ(TBMS Ⅱ)on the death of colorectal cancer cells LOVO by promoting the degradation of nuclear erythroid factor 2-like(NRF2),the cell viability assay was performed to detect the inhibitory effect of TBMS Ⅱ on LOVO cells.Human colorectal cancer LOVO cells were cultured,and the inhibitory effect of TBMS Ⅱ on LOVO cells was detected by cell viability assay,the IC_(50) value of half lethality was calculated,and the cells were grouped according to the IC_(50) value.The effect of the clone formation ability of the cells was detected by clone formation assay.The effect of apoptosis,cell cycle,and the level of ROS in the cells was detected by flow cytometry.And the effect of TBMS Ⅱ on The effects of TBMS Ⅱ on the levels of reduced glutathione(GSH),superoxide dismutase(SOD),total antioxidant capacity(T-AOC)and malondialdehyde(MDA)were detected by biochemical kits.The effects of TBMS Ⅱ on the expression of NRF2 protein in LOVO cells were detected by Western blot assay.Molecular docking was performed to investigate the binding capacity between TBMS Ⅱ and NRF2.The results showed that TBMS Ⅱ significantly inhibited the viability,clone formation ability,affected the cell cycle,and induced apoptosis in LOVO cells.TBMS Ⅱ inhibited the levels of antioxidant indexes GSH,SOD,and T-AOC,and up-regulated the levels of MDA,and ROS in the cells.TBMS Ⅱ inhibited the expression of NRF2protein,and up-regulated MDA,and ROS in the cells,probably by directly binding to NRF2.TBMS Ⅱ may directly bind to NRF2,inhibit the expression of NRF2 protein in cells,and promote the degradation of NRF2 protein in cells.The inhibition of NRF2 expression enhances the apoptosis-inducing ability of TBMS Ⅱ.Therefore,TBMS Ⅱ induced apoptosis in colorectal cancer LOVO cells by promoting the degradation of NRF2 protein.

关 键 词：土贝母苷乙 NRF2 蛋白降解 结直肠癌细胞LOVO 

分 类 号：Q28[生物学—细胞生物学]