5-LOX抑制剂NDGA对人肝癌HepG-2细胞裸鼠移植瘤生长的影响及其机制的探讨  

Investigate the effect and possible mechanism of 5-lipoxygenase (5-LOX) inhibitor NDGA on proliferation of HepG2 cell Xenografts in small nude mice

作　　者：张雪兵[1] 俞广进[1] 夏俊[1] 贾冉[1] 陈双升 熊奇如[1] 

机构地区：[1]安徽医科大学第一附属医院肝胆外科,合肥230022

出　　处：《肝胆外科杂志》2014年第5期380-383,共4页Journal of Hepatobiliary Surgery

摘　　要：目的观察5-lox抑制剂去甲二氢愈创木酸(NDGA)对裸鼠人肝癌HepG-2细胞移植瘤的作用,并探讨其抗肿瘤的可能机制。方法以人肝癌细胞HepG-2制备裸鼠人肝癌移植瘤模型共18只,将18只荷瘤裸鼠随机分为三组:1、正常对照组(接种正常肝癌HepG2细胞)。2、药物组(接种正常肝癌HepG2细胞同时皮下注入5-lox抑制剂去甲二氢愈创木酸(NDGA),剂量:10μml/L NDGA 0.1ml/10g一天一次,连续5天)。3、溶媒组(接种正常HepG-2细胞后皮下注射溶质二甲亚酚)。密切观察移植瘤生长及裸鼠生存情况21天,记录各组移植瘤肿瘤体积V(V=长径×短径2×0.5),计算衰退率:肿瘤衰退率=1-干预组平均肿瘤体积比/空白对照组平均肿瘤体积比(V/Vo)。通过RT-PCR及Western-blot法检测裸鼠移植瘤组织中bcl-2、caspase3凋亡调节因子以及通路信号蛋白MEK1/2、ERK1/2等的表达。结果实验期间各组荷瘤鼠活动较好,成瘤率100%,且实验过程中无不良繁衍及裸鼠死亡;试验后分别测得各组荷瘤鼠皮下移植瘤体积及肿瘤衰退率,利用单因素方差分析及t检验可以发现:药物组荷瘤鼠较对照组及溶媒组移植瘤体积明显缩小,肿瘤衰退率明显较高,且存在统计学差异(P<0.01);利用RT-PCR及Western-blot法检测发现药物组caspase3表达量较对照组及溶媒组明显增强,差异存在统计学意义(P<0.01),bcl-2、ERK1/2、MEK1/2等表达量较对照组及溶媒组明显下降,差异具有统计学意义(P<0.01)。结论 NDGA对于裸鼠人肝癌移植瘤具有明显的抑制效果。NDGA可能通过诱导肿瘤细胞凋亡而抑制肿瘤生长,其作用机制与抑制MEK/ERK信号通路有关。Objective To investigate the effect of the5-lipoxygenase（ 5-LOX） inhibitor,NDGA on human hepatocellular carcinoma xenografts in vivo and to explore its potential anti- neoplasm mechanism. Methods Cultured HepG-2 human hepatocellular carcinoma cells were injected into the flanks of eighteen nude mice to develop xenograft models and the nude mice were randomized into three groups to be administered with NDGA dissolved into DMSO,DMSO only,nothing,respectively. The nude mice were monitored for 21 consecutive days for tumor volume changes. Then the tumor tissues were isolated and recorded each group xenograft tumor volume（ V = ab2 /2mm3）,the tumor inhibition rate（ V /V0＊ 100%）. RT-PCR and Western blot were used to detect the mRNA and protein expression of apoptosis regulatory factor of xenograft tumor tissue（ bcl-2,caspase3）,and pathway signaling proteins（ MEK1 /2,ERK1 /2）. Results None of the 18 nude mice died during the experiment and all mice formed hepatocellular carcinoma xenografts in situ. NDGA inhibited growth of human HepG-2 hepatocellular carcinoma xenografts in nude mice,measured as both tumor volume and tumor inhibition rate. By RT-PCR and Western-blot assay was found caspase3 in the drug group was significantly increased compared with the control group and the vehicle group,while the bcl-2、ERK1 /2、MEK1 /2 was significantly decreased（ P〈0. 01）. Conclusions5- LOX inhibitor,NDGA,inhibits the growth of hepatocellular carcinoma xenografts by inducing the apoptosis of human hepatocellular carcinoma cells. The mechanism could be related to inhibition of MEK /ERK signaling pathway.

关 键 词：脂氧合酶抑制剂 肝癌 细胞凋亡 细胞外信号调节激酶 

分 类 号：R735.7[医药卫生—肿瘤]