五倍子酸对肝癌HepG2细胞索拉非尼化疗增敏作用  

Effect of gallic acid in increasing the chemosensitivity of hepatocellular carcinoma HepG2 cells to sorafenib

作　　者：刘百坤[1] 王志茹[1] 赵文静[1] LIU Baikun;WANG Zhiru;ZHAO Wenjing(Central Laboratory of Hepatobiliary Hospital of Jilin,Changchun 130062,China)

机构地区：[1]吉林省肝胆病医院中心实验室,长春130062

出　　处：《临床肝胆病杂志》2025年第2期292-299,共8页Journal of Clinical Hepatology

基　　金：吉林省卫生与健康技术创新项目(2020J082)。

摘　　要：目的观察五倍子酸(GA)联合索拉非尼(Sora)对HepG2细胞的化疗增敏作用,并探讨其作用机制。方法将肝癌HepG2细胞分为对照组、GA组、Sora组和GA+Sora组。CCK8法检测细胞活力,CompuSyn软件分析联合用药指数(CI值);平板克隆形成实验检测细胞克隆形成能力;流式细胞术检测细胞凋亡;细胞划痕和Transwell小室侵袭实验检测细胞迁移和侵袭能力;Western Blot法检测基质金属蛋白酶(MMP)2、MMP-9和凋亡相关蛋白表达。肝癌HepG2细胞接种于裸鼠背部右下方,植瘤6 d后分为4组:对照组(Control)、GA组、Sora组和GA+Sora组,每周测量1次肿瘤大小和质量,药物干预21 d,处死裸鼠,取肿瘤称重。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果GA和Sora作用HepG2细胞48 h的IC_(50)值分别为(123.47±5.16)μmol/L和(9.87±0.98)μmol/L,Sora与70μmol/L GA(IC_(30))联用后,IC_(50)降至(2.06±0.35)μmol/L,不同浓度Sora与70μmol/L GA联合用药CI值均<1;各组细胞克隆形成数分别为234.0±20.4、147.0±12.1、129.3±13.3、73.0±7.6,GA+Sora组细胞克隆数明显低于对照组、GA组和Sora组(P值均<0.05);处理48 h后,各组细胞凋亡率分别为(1.98±0.29)%、(15.17±1.56)%、(18.65±1.48)%和(34.60±5.36)%,GA+Sora组细胞凋亡率明显高于对照组、GA组和Sora组(P值均<0.05);处理24 h后,各组细胞迁移率分别为(55.59±5.08)%、(29.34±4.36)%、(21.80±5.16)%和(6.47±2.75)%,GA+Sora组细胞迁移率明显低于对照组、GA组和Sora组(P值均<0.05);处理48 h后,各组穿膜细胞数分别为223.7±13.0、168.3±10.9、155.3±29.1和62.7±19.7,GA+Sora组穿膜细胞数低于对照组、GA组和Sora组(P值均<0.05);与对照组比较,GA组、Sora组和GA+Sora组细胞中MMP-2、MMP-9、B淋巴细胞瘤2(Bcl-2)蛋白表达水平均明显降低(P值均<0.05),Bcl-2关联X蛋白(Bax)、裂解的半胱氨酸天冬氨酸蛋白酶3(Cleaved caspase-3)蛋白表达水平均明显升高(P值均<0.05)。GA组、SoObjective To investigate the chemosensitization effect of gallic acid(GA)combined with sorafenib(Sora)on hepatocellular carcinoma HepG2 cells and related mechanisms.Methods HepG2 cells were randomly divided into control group,GA group,Sora group,and GA+Sora group.CCK8 assay was used to measure cell viability;CompuSyn software was used to analyze combination index(CI);colony formation assay was used to evaluate the colony formation ability of cells;flow cytometry was used to measure cell apoptosis;wound healing assay and Transwell chamber assay were used to observe the migration and invasion abilities of cells;Western Blot was used to measure the expression matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9),and apoptosis-related proteins.HepG2 cells were subcutaneously inoculated into the lower right back of mice,and 6 days later,the mice were divided into control group,GA group,Sora group,and GA+Sora group.Tumor size and body weight were measured once a week,and drug intervention was performed for 21 days.Then the nude mice were sacrificed,and tumor weight was measured.A one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results The mean IC_(50) values of GA and Sora for the treatment of HepG2 cells for 48 hours were 123.47±5.16μmol/L and 9.87±0.98μmol/L,respectively,and when Sora was combined with 70μmol/L GA(IC_(30)),IC_(50) decreased to 2.06±0.35μmol/L;the CI value was<1 for Sora at different concentrations combined with 70μmol/L GA.The number of cell colonies was 234.0±20.4,147.0±12.1,129.3±13.3,and 73.0±7.6,respectively,in the four groups,and the GA+Sora group had a significantly lower number of cell colonies than the control group,the GA group,and the Sora group(all P<0.05).After 48 hours of treatment,the cell apoptosis rate was 1.98%±0.29%,15.17%±1.56%,18.65%±1.48%,and 34.60%±5.36%,respectively,in the four groups,and the GA+Sora group had a significantly

关 键 词：癌 肝细胞 索拉非尼 五倍子酸 

分 类 号：R285.5[医药卫生—中药学]