盐酸川芎嗪对人肝癌细胞及外泌体蛋白GPC3的影响  

Effect of tetramethylpyrazine hydrochloride on hepatocellular carcinoma cells and exosome protein GPC3

作　　者：刘振华[1] 王宁 李克跃[1] 汤可立[1] 石承先[1] 丁杰[3] 王飞清 LIU Zhenhua;WANG Ning;LI Keyue;TANG Keli;SHI Chengxian;DING Jie;WANG Feiqing(Department of Hepatobiliary Surgery,Guizhou Provincial People′s Hospital,Guiyang, Guizhou 550002,China;Department of Pharmacy,Guizhou Provincial Orthopedic Hospital, Guiyang,Guizhou 550004,China;Department of Gastrointestinal Surgery,Guizhou Provincial People′s Hospital,Guiyang,Guizhou 550002,China;Department of Clinical Laboratory, First Affiliated Hospital of Guiyang College of Traditional Chinese Medicine,Guiyang,Guizhou 550001,China)

机构地区：[1]贵州省人民医院肝胆外科,贵阳550002 [2]贵州省骨科医院药剂科,贵阳550004 [3]贵州省人民医院胃肠外科,贵阳550002 [4]贵阳中医学院附属第一医院检验科,贵阳550001

出　　处：《重庆医学》2018年第33期4226-4228,4233,共4页Chongqing medicine

基　　金：贵州省中医药;民族医药科学技术研究课题资助(GZYY-2015-002)

摘　　要：目的探讨盐酸川芎嗪(TMPH)对肝癌细胞外泌体蛋白磷脂酰肌醇蛋白聚糖-3(GPC3)的影响,以了解其抑制肝癌的机制。方法采用Western blot分别检测2种肝癌细胞(MHCC97-H、HepG2)及TMPH处理24h后上清液外泌体GPC3的表达水平。利用共培养法培养HepG2细胞,TMPH处理24h后采用划痕实验和Transwell实验分析与MHCC97-H共培养的HepG2细胞侵袭迁移能力。结果与HepG2相比,MHCC97-H上清液外泌体蛋白GPC3水平升高,表达差异有统计学意义(P<0.01),加入TMPH处理MHCC97-H 24h后,上清液中外泌体蛋白GPC3水平明显降低(P<0.01);与MHCC97-H共培养的HepG2侵袭迁移能力明显增强,GPC3水平较高,差异均有统计学意义(P<0.01);TMPH处理24h后HepG2侵袭迁移能力减弱,且GPC3水平明显下降(P<0.01)。结论 TMPH调节外泌体中GPC3能对HepG2的侵袭迁移产生作用,这可能是其抗肝癌的机制之一。Objective To investigate the effect of tetramethylpyrazine hydrochloride(TMPH)on exosome protein glypican-3(GPC3)of hepatocellular carcinoma(HCC)cells for understanding its mechanism for inhibiting HCC.Methods Western blot was used to detect the expression level of GPC3 in supernatant of two kinds of hepatoma cells(MHCC97-H,HepG2)and TMPH treatment for 24 h respectively.HeG2 cells were cultured by using the co-culture method.After TMPH treatment for 24 h,the scratch test and Transwell assay were used to analyze the invasion and migration of HepG2 cells co-cultured with MHCC97-H.Results Compared with HepG2,the level of exosome protein GPC3 in supernatant of MHCC97-H was significantly higher,the expression difference was statistically significant(P<0.01),and the expression of GPC3 in supernatant was significantly decreased after adding TMPH to treat MHCC97-H for 24 h(P<0.01);the invasion and migration ability of HepG2 cells co-cultured with MHCC97-H was increased significantly,and the level of GPC3 was higher(P<0.01).The invasion and migration ability of HepG2 cells decreased after TMPH treatment for 24 h,moreover the GPC3 level was decreased significantly(P<0.01).Conclusion TMPH regulating GPC3 in exosomes produce the effect on the invasion and migration of HepG2,which may be one of the mechanisms of its anti-liver cancer.

关 键 词：川芎嗪 肝癌细胞 磷脂酰肌醇蛋白聚糖-3 

分 类 号：R735.7[医药卫生—肿瘤]