罗哌卡因对人结肠癌顺铂耐药细胞株顺铂敏感性的增强作用及其机制  

Ropivacaine enhances cisplatin sensitivity of human colorectal cancer cisplatin-resistant cell lines and its mechanism

作　　者：赵文博 曾炼 胡鹏超 程欣冉 罗辉宇 ZHAO Wenbo;ZENG Lian;HU Pengchao;CHENG Xinran;LUO Huiyu(Department of Anesthesiology,Xiangyang No.1 People's Hospital Affiliated to Hubei University of Medicine,Xiangyang 441000,China)

机构地区：[1]湖北医药学院附属襄阳市第一人民医院麻醉科,湖北襄阳441000 [2]湖北医药学院基础医学院

出　　处：《山东医药》2023年第27期21-28,共8页Shandong Medical Journal

摘　　要：目的观察罗哌卡因对人结肠癌顺铂耐药细胞株顺铂敏感性的增强作用,并进一步探讨其作用机制。方法通过浓度递增法将人结肠癌LOVO细胞暴露于顺铂中诱导顺铂耐药株LOVO/DDP形成。将LOVO和LOVO/DDP两种结肠癌细胞株分别分为罗哌卡因联合顺铂组、罗哌卡因组、顺铂组、对照组。对照组加入细胞培养液,顺铂组加入细胞培养液和1.0μmol/L浓度顺铂,罗哌卡因联合顺铂组加入细胞培养液后再同时加入1.0μmol/L浓度顺铂及1 mmol/L浓度罗哌卡因培养。培养14 d,采用平板克隆实验观察细胞增殖能力。培养48 h后,采用Tran⁃swell和流式细胞术观察细胞迁移、凋亡情况;JC-1和ROS实验测定细胞线粒体膜电位和活性氧(ROS);Western blot⁃ting法、免疫荧光法检测细胞株中MMP-9及铁死亡相关蛋白GPX4、Nrf-2、SLC7A11;亚铁离子染色法检测结肠癌细胞株中亚铁离子。结果LOVO细胞中,与对照组比较,顺铂组LOVO细胞株克隆体的形成减少、迁移细胞株数减少、MMP-9蛋白水平降低、凋亡率升高、存活率降低,罗哌卡因组克隆体形成数、迁移细胞株数、MMP-9蛋白水平、细胞凋亡率、细胞存活率变化不明显,顺铂联合罗哌卡因组细胞株克隆体数减少、迁移细胞数减少、MMP-9蛋白水平下降、凋亡率升高、存活率降低(P均<0.05);与顺铂组比较,顺铂联合罗哌卡因组细胞株克隆体数少、迁移细胞数少、MMP-9蛋白水平低、凋亡率高、存活率低(P均<0.05)。在LOVO/DDP细胞株中,与对照组比较,顺铂组、罗哌卡因组细胞株克隆体数、迁移细胞株数、MMP-9蛋白水平变化不明显,顺铂联合罗哌卡因组细胞株克隆体数、迁移细胞株数减少、MMP-9蛋白水平降低、细胞凋亡率降低、细胞存活率升高(P均<0.05)。在LOVO细胞株和LOVO/DDP细胞株中,与对照组比较,其他三组JC-1多聚体红色荧光与单体绿色荧光比值[JC-1(Red/Green)]均降低,ROS水平升高,GPXObjective To observe the enhancement effect of ropivacaine on cisplatin sensitivity of human colorectal cancer cells and further to explore its mechanism of action.Methods The human colorectal cancer LOVO cells were ex⁃posed to cisplatin through a concentration-increasing method to induce the formation of cisplatin-resistant strain LOVO/DDP.The colorectal cancer cell lines LOVO and LOVO/DDP were divided into the ropivacaine combined with cisplatin group,ropivacaine group,cisplatin group,and control group,respectively.Cells in the control group were added with cell culture fluid,cells in the ropivacaine group were added with cell culture fluid and 1 mmol/L ropivacaine,cells in the cispl⁃atin group were added with cell culture fluid and 1.0μmol/L cisplatin,and cells in the ropivacaine combined with cisplat⁃in group were first added with cell culture fluid and then were added with 1.0μmol/L cisplatin and 1 mmol/L ropiva⁃caine.After 14-day culture,the cell proliferation ability was observed by plate cloning assay.After 48-hour culture,Tran⁃swell and flow cytometry were used to observe the cell migration and apoptosis;JC-1 and ROS experiments were used to de⁃termine cell mitochondrial membrane potential and reactive oxygen species(ROS);Western blotting and immunofluores⁃cence were used to detect matrix metalloproteinase-9(MMP-9)and ferroptosis-related proteins glutathione peroxidase 4(GPX4),nuclear erythroid 2-related factor 2(Nrf-2),and solute carrier family 7 member 11(SLC7A11)in the cell lines;ferroptosis staining method was used to detect ferrous iron ions in colorectal cancer cell lines.Results In LOVO cells,compared with the control group,the formation of clones of the LOVO cell line in the cisplatin group decreased,the number of migrating cell lines decreased,the MMP-9 protein level was reduced,the apoptosis rate increased,and the sur⁃vival rate was reduced in the cisplatin group;the ropivacaine group had no significant changes in formation of clones,cell migration number,MMP-9 protein level,a

关 键 词：罗哌卡因 人结肠癌细胞株 耐药细胞株 铁死亡 核红细胞2相关因子2/谷胱甘肽过氧化物酶4信号通路 

分 类 号：R641[医药卫生—外科学]