摘要
目的探究苦参碱通过介导Janus激酶2/信号转导和转录激活因子3(JAK2/STAT3)信号通路对肝癌细胞迁移、侵袭和上皮间质转化(EMT)的影响。方法将体外培养人肝癌细胞MHCC97-H分为:对照组(不做处理),苦参碱组(0.125、0.25、0.5、1和2 mg/mL苦参碱),AG490组(10μmol/L JAK2/STAT3信号通路抑制剂AG490),抑制剂组(0.5 mg/mL苦参碱和10μmol/L AG490),干预24 h。MHCC97-H细胞活力采用细胞计数试剂盒法测定;细胞迁移和侵袭能力采用Transwell小室法测定;EMT相关因子mRNA表达采用RT-qPCR法测定;蛋白水平采用蛋白免疫印迹法测定。结果与对照组相比,0.125、0.25和0.5 mg/mL苦参碱组细胞活力无显著影响(P>0.05),1和2 mg/mL苦参碱组和AG490组细胞活力显著降低(P<0.05),其中0.5 mg/mL苦参碱组MHCC97-H细胞迁移数、侵袭数和p-JAK2/JAK2、p-STAT3/STAT3、N-钙粘蛋白、波形蛋白及纤连蛋白表达水平显著降低(P<0.05),E-钙粘蛋白表达水平显著增加(P<0.05),加入JAK2/STAT3信号通路抑制剂后,抑制剂组显著加剧了苦参碱抑制细胞迁移、侵袭和EMT的能力(P<0.05)。结论苦参碱可能是通过调控JAK2/STAT3通路的信号转导来抑制MHCC97-H细胞的迁移、侵袭和EMT。
Objective To explore the effect of matrine on the migration,invasion and EMT of hepatocellular carcinoma cells by mediating the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway.Methods Human hepatocellular carcinoma MHCC97-H cells were cultured in vitro,and divided into:control group(no treatment),matrine groups(0.125,0.25,0.50,1 and 2 mg/mL matrine),AG490 group(10μmol/L JAK2/STAT3 signaling inhibitor AG490)and inhibitor group,treated for 24 hours.The MHCC97-H cell viability was determined by cell counting kit-8 method.The cell migration and invasion abilities were determined by Transwell assay.EMT-related factors mRNA expression was determined by RT-qPCR.Relative expression levels were determined by western blot.Results Compared with the control group,the 0.125,0.25 and 0.5 mg/mL matrine groups had no significant effect on the cell viability(P>0.05),while the 1 and 2 mg/mL matrine groups and AG490 group had a significant reduction in the cell viability(P<0.05).The 0.5 mg/mL matrine group significantly decreased the number of cell migration and invasion and expression levels of p-JAK2/JAK2,p-STAT3/STAT3,N-cadherin,Vimentin and fibronectin(P<0.05),and significantly increased E-cadherin expression level(P<0.05).After adding JAK2/STAT3 signaling pathway inhibitor,inhibitor group significantly intensified the ability of matrine to inhibit cell migration,invasion and EMT(P<0.05).Conclusion Matrine significantly inhibited the migration,invasion and EMT of MHCC97-H cells.Its mechanism may be related to the inactivation of JAK2/STAT3 signling pathway.
作者
朱丽
胡朝霞
胡瑞晴
ZHU Li;HU Hai-xia;HU Rui-qing(Second Department of Liver Disease,Chengdu Public Health Clinical Medical Center,Chengdu,Sichuan,610000,China)
出处
《现代消化及介入诊疗》
2023年第8期976-980,共5页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
四川省科技计划项目(2020YFS0485)。
关键词
苦参碱
肝癌
迁移
侵袭
上皮间质转化
Matrine
Hepatocellular carcinoma
Migration
Invasion
Epithelial-mesenchymal transition
