石榴汁含样血清对肝癌细胞JHH-7凋亡和迁移的影响及机制初探  

Effects of pomegranate juice-containing serum on apoptosis and migration of hepatocellular carcinoma JHH-7 cells and its mechanism

作　　者：田丽 康宇鸿 周婷[1] 黄孝懿 夏锐 邓远乐 王礼群 李亚丽[1] 阴文娅[1] TIAN Li;KANG Yu-hong;ZHOU Ting;HUANG Xiao-yi;XIA Rui;DENG Yuan-le;WANG Li-qun;LI Ya-li;YIN Wen-ya(West China School of Public Health and West China Fourth Hospital,Sichuan University,Chengdu,Sichuan 610041,China)

机构地区：[1]四川大学华西公共卫生学院/四川大学华西第四医院,四川成都610041

出　　处：《现代预防医学》2022年第5期883-887,926,共6页Modern Preventive Medicine

摘　　要：目的研究石榴汁含样血清对肝癌细胞JHH-7凋亡与迁移的影响,并探讨其可能的机制。方法雄性SD大鼠随机分为石榴汁组(灌胃石榴汁20 mL/(kg·bw))和空白组(灌胃等量生理盐水),连续灌胃7 d,2次/d。末次灌胃后腹主动脉采血,离心获得含样和空白血清,分别干预肝癌细胞JHH-7,并以胎牛血清作为对照。MTT法检测细胞活性,计算细胞存活率;Annexin V-FITC/PI双染法检测细胞的凋亡情况,Western blot检测线粒体凋亡途径相关蛋白Bcl-2、Bax、p53、Caspase-3、Cytochrome-c(Cyt-C)和Caspase-9以及迁移相关蛋白MMP2、MMP9和TIMP2的表达;划痕实验检测细胞的迁移能力。结果与空白血清相比,含样血清可抑制JHH-7细胞的增殖,具有时间依赖性,且空白血清和含样血清对人正常肝细胞LO;未表现出抑制作用;与空白血清相比,含样血清干预JHH-7细胞后,细胞凋亡率显著增加(P<0.05),激活了Bcl-2家族蛋白,进一步导致线粒体损伤,诱发了线粒体途径诱导的细胞凋亡,相关蛋白p53、Caspase-3、Cyt-C和Caspase-9表达上调,Bcl-2的表达下调,Bax/Bcl-2比值增加(P<0.05);与空白血清相比,含样血清抑制了JHH-7细胞的迁移,且相关蛋白MMP2和MMP9的表达下调,TIMP2的表达上调,TIMP2/MMP2比值增加(P<0.05)。结论石榴汁含样血清通过线粒体途经诱导肝癌细胞JHH-7的凋亡,并且通过调节MMP2/TIMP2和MMP9表达的平衡抑制JHH-7的迁移,从而发挥抗肝癌作用。Objective To investigate the effects of pomegranate juice-containing serum on apoptosis and migration of hepatoma cell line JHH-7,and to explore its possible mechanism.Methods Male SD rats were randomly divided into a pomegranate juice group(gavage with pomegranate juice 20 mL/(kg·bw))and a blank group(gavage with the same amount of normal saline).The samples were continuously administered for 7 days,2 times/d.After the last gavage,rat serum was taken as the intervention sample,and pomegranate juice-containing serum and blank serum were used to intervene liver cancer cells JHH-7,respectively.Fetal bovine serum was used as a control.MTT assay was performed to detect the effect of sample serum on the survival rate of JHH-7 cells,and the cell survival rate was calculated.Cell apoptosis was detected by Annexin V-FITC/PI double staining.The expressions of mitochondrial apoptosis pathway related proteins Bcl-2,Bax,p53,Caspase-3,Cytochrome-C(Cyt-C)and Caspase-9,as well as migration-related proteins MMP2,MMP9 and TIMP2 were detected by Western blot.The wound healing assay was adopted for the migration ability of cells.Results Compared with the blank serum,the serum containing samples could inhibit the proliferation of JHH-7 cells in a time dependent manner,and the blank serum and the serum containing samples showed no inhibitory effect on human normal hepatocyte LO;.Compared with the blank serum,the apoptosis rate of JHH-7 cells was significantly increased after the intervention of serum containing samples(P<0.05),which had activated Bcl-2 family proteins,further caused mitochondrial damage and induced cell apoptosis by mitochondrial pathway.The expression of p53,Caspase-3,Cyt-C and Caspase-9 were up-regulated,the expression of Bcl-2 was down-regulated,and the ratio of Bax/Bcl-2 was increased(P<0.05).Compared with the blank serum,the serum containing samples inhibited the migration of JHH-7 cells,and the expressions of MMP2 and MMP9 were down-regulated,the expression of TIMP2 was up-regulated,and the TIMP2/MMP2 ratio wa

关 键 词：石榴汁含样血清 JHH-7 细胞凋亡 细胞迁移 线粒体途径 

分 类 号：R151.2[医药卫生—营养与食品卫生学]