甲基纳曲酮对肝癌HepG2细胞体内外增殖及体外迁移能力的影响

Effects of methylnaltrexone on in vitro and vivo proliferation and in vitro migration of hepatoma HepG2 cells

目的探讨甲基纳曲酮对肝癌Hep G2细胞体内外增殖及体外迁移能力的影响。方法 (1)将Hep G2细胞分为低、高浓度甲基纳曲酮组及空白对照组,低、高浓度甲基纳曲酮组分别给予终浓度为100μmol/L、1 000μmol/L的甲基纳曲干预,空白对照组给予等体积生理盐水,分别于培养24 h、48 h、72 h后检测细胞增殖能力,72 h后检测细胞迁移能力,并检测高浓度甲基纳曲酮组及空白对照组Hep G2细胞中血管内皮生长因子(VEGF)和基质金属蛋白酶(MMP)9蛋白的表达水平。(2)将0.2 ml Hep G2细胞(浓度为1×107/ml)接种于12只BALB/C裸鼠腋下,接种第8天后,将裸鼠随机分为对照组和甲基纳曲酮组各6只,甲基纳曲酮组腹腔注射甲基纳曲酮,对照组给予等量生理盐水,连续给药21 d,比较两组体重及净瘤质量。结果高、低浓度甲基纳曲酮组细胞增殖抑制率均高于空白对照组,且均随时间延长而升高,迁移细胞数均少于空白对照组(P<0.05),仅高浓度甲基纳曲酮组VEGF、MMP9蛋白的表达水平均低于对照组(P<0.05);甲基纳曲酮组裸鼠体重大于对照组,而净瘤质量低于对照组(P<0.05)。结论甲基纳曲酮可以抑制肝癌Hep G2细胞体内外增殖及体外迁移,其机制可能与下调VEGF、MMP9的表达有关。

Objective To explore the effects of methylnaltrexone on the in vitro and vivo proliferation and in vitro migration of hepatoma HepG2 cells. Methods （1）The HepG2 cells were divided into low-dose methylnaltrexone group,high-dose methylnaltrexone group and blank control group. The low- and high-dose methylnaltrexone groups were given 100 μmol/L methylnaltrexone and 1 000 μmol/L methylnaltrexone,respectively,and the blank control group received the equivalent volume of normal saline.The cell proliferation was detected after 24,48 and 72 hours of intervention,then after 72 hours of intervention,the cell migration was determined,and the expression levels of vascular endothelial growth factor（VEGF） and matrix metalloproteinase（MMP） 9 proteins in HepG2 cells were detected in the high-dose methylnaltrexone group and blank control group.（2）Subaxillary incubation with 0.2 ml HepG2 cells（1×10 7/ml） was performed in 12 BALB/C nude mice,then on the 8th day after incubation,the mice were randomly divided into control group and methylnaltrexone group,with 6 mice in each group.The methylnaltrexone group was given intraperitoneal injection of methylnaltrexone,while the control group was given the equivalent volume of normal saline,the intervention lasted for 21 days in both groups,then the body weight and net tumor mass of mice were compared between the two groups. Results In the low- and high-dose methylnaltrexone groups,the proliferation inhibition rate was higher,which increased over time,and the number of migration cells was lower than those in the blank control group（ P 〈0.05）.The expression levels of VEGF and MMP9 proteins in the high-dose methylnaltrexone group were lower than those in the blank control group （ P 〈0.05） ; the body weight was greater while the net tumor mass was lower in the methylnaltrexone groups than those in the control group（ P 〈0.05）. Conclusion Methylnaltrexone can inhibit the in vitro and vivo proliferation and in vitro migration of hepatoma HepG2 cells,which might be related to down-regulation of VEGF and MMP9.