麝香酮对胰腺癌中NLRP3炎症体通路介导的上皮间质转化的影响  

Effect of muscone on NLRP3 inflammatory body pathway-mediated epithelial-mesenchymal transformation in pancreatic cancer

作　　者：宋银雪[1] 苗常青[1] 许静[1] 杜俊凯[1] 徐之超[1] SONG Yinxue;MIAO Changqing;XU Jing;DU Junkai;XU Zhichao(Department of Emergency,The First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,China)

机构地区：[1]西安交通大学第一附属医院急诊科,陕西西安710061

出　　处：《西部医学》2024年第5期642-648,共7页Medical Journal of West China

基　　金：陕西省重点研发计划项目(2021SF-234)。

摘　　要：目的探讨麝香酮(MUS)对胰腺癌(PC)的抗癌作用及其对核苷酸结合寡化结构域样受体3(NLRP3)炎症体通路和上皮间质转化(EMT)的影响。方法根据MUS处理浓度不同将人胰腺癌细胞(PANC1)细胞分为0μM组、0.1μM组、0.5μM组、1μM组、2μM组和4μM组,各组细胞使用相应浓度的MUS孵育48 h。使用四甲基偶氮唑盐(MTT)法测定生长抑制率和半抑制浓度(IC50)值,使用Annexin V-FITC/PI染色法测定细胞凋亡,使用Transwell测定细胞迁移和侵袭。使用Western blot测定Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤(Bcl-2)、基质金属蛋白酶(MMP2)、MMP9、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、NLRP3、半胱氨酸蛋白酶-1(caspase-1)和血清人细胞凋亡相关斑点样蛋白(ASC)的蛋白表达水平。通过对BALB/c雄性裸鼠皮下注射PANC1细胞构建移植瘤模型,然后将裸鼠随机分为对照组和MUS组,每组6只。对照组裸鼠给予腹腔注射100μL的1%二甲基亚砜(DMSO),MUS组裸鼠给予腹腔注射100μL的MUS溶液(20 mg/kg)。给药21 d后测量裸鼠肿瘤体积和重量。结果与0μM组相比,0.1μM组、0.5μM组、1μM组、2μM组和4μM组的PANC1细胞的生长抑制率逐渐升高,凋亡率逐渐升高(均P<0.05),Bax蛋白表达水平逐渐升高,Bcl-2蛋白表达水平逐渐降低(P<0.05)。与0μM组相比,0.1μM组、0.5μM组、1μM组、2μM组和4μM组的迁移和侵袭细胞数逐渐降低(P<0.05),MMP2和MMP9蛋白表达水平逐渐降低(P<0.05)。与0μM组相比,0.1μM组、0.5μM组、1μM组、2μM组和4μM组的E-cadherin蛋白表达水平逐渐升高,N-cadherin和Vimentin蛋白表达水平逐渐降低(P<0.05),NLRP3、caspase-1和ASC蛋白表达水平逐渐降低(P<0.05)。与对照组比较,MUS组裸鼠的肿瘤体积和重量均降低(P<0.05),肿瘤组织中的NLRP3、caspase-1和ASC蛋白表达水平均降低(P<0.05)。结论MUS具有良好的抗PC作用,可能通过抑制NLRP3炎症体通路介导的EMT发挥抗癌作用。Objective To investigate the anticancer effect of muscone(MUS)on pancreatic cancer and its effect on NLRP3 inflammatory body pathway and epithelial-mesenchymal transformation(EMT).Methods PANC1 cells were divided into 0μM group,0.1μM group,0.5μM group,1μM group,2μM group and 4μM group according to the different concentration of muscone treatment.The growth inhibition rate and half maximal inhibitory concentration(IC50)were measured by MTT method,apoptosis was detected by Annexin V-FITC/PI staining,and cell migration and invasion were measured by Transwell.The protein expression levels of Bax,Bcl-2,MMP2,MMP9,E-cadherin,N-cadherin,Vimentin,NLRP3,caspase-1 and ASC were measured by Western blot.The transplanted tumor model was established by subcutaneous injection of PANC1 cells into BALB/c male nude mice,and then the nude mice were randomly divided into control group and MUS group(n=6).The nude mice in control group were injected intraperitoneally with 100μL 1%DMSO,and the nude mice in MUS group were injected intraperitoneally with 100μL MUS solution(20 mg/kg).The volume and weight of tumor were measured 21 days after administration.Results Compared with 0μM group,the growth inhibition rate and apoptosis rate of PANC1 cells in 0.1μM group,0.5μM group,1μM group,2μM group and 4μM group increased gradually,and the expression level of Bax protein increased gradually,while the expression level of Bcl-2 protein decreased gradually(P<0.05).Compared with 0μM group,the number of migration and invasion cells in 0.1μM group,0.5μM group,1μM group,2μM group and 4μM group decreased gradually,and the expression of MMP2 and MMP9 protein decreased gradually(P<0.05).Compared with 0μM group,the expression level of E-cadherin protein in 0.1μM group,0.5μM group,1μM group,2μM group and 4μM group increased gradually,while the expression level of N-cadherin and Vimentin protein decreased gradually,and the expression level of NLRP3,caspase-1 and ASC protein decreased gradually(P<0.05).Compared with control group,th

关 键 词：麝香酮 胰腺癌 肿瘤转移 NLRP3炎症体通路 上皮间质转化 

分 类 号：R735.9[医药卫生—肿瘤]