过氧化物酶1对人肝癌细胞HepG2细胞皮下移植瘤生长的影响  

Effect of Prx-1 on growth of hepatocellular carcinoma xenografts

作　　者：涂青松[1] 何剪太[1] 

机构地区：[1]中南大学湘雅医院,湖南长沙410008

出　　处：《中国现代医学杂志》2016年第8期11-16,共6页China Journal of Modern Medicine

摘　　要：目的探讨过氧化物酶1(Prx 1)在肝癌形成和恶性进展中的作用。方法蛋白质印迹法(Western blot)检测远癌肝组织和近癌肝组织中Prx 1、p Akt及Akt表达差异。肝细胞饥饿培养24 h后,10 ng/ml表皮生长因子(EGF)处理10 min,提取总蛋白,Western blot检测Prx 1、p Akt、不同位点磷酸化表皮生长因子受体(EGFR)和总EGFR。Western blot检测检测未处理组、转染无义sh RNA组及转染Prx 1 sh RNA组Hep G2细胞中Prx 1蛋白的表达。DCFH-DA法检测3种Hep G2细胞中活性氧簇总量。转染无义sh RNA组及转染Prx 1sh RNA组Hep G2细胞皮下注射至SCID小鼠,注射后每周测量1次皮下肿瘤体积。Western blot检测两组小鼠皮下肿瘤组织中Prx 1和p Akt的表达情况。结果与正常肝细胞比较,Prx 1在肝转化细胞中表达升高。Prx1增强了EGF介导的EGFR和Akt激活。Prx 1表达沉默抑制了Hep G2体内成瘤能力。减少Hep G2皮下移植瘤的肝转移。Prx 1表达沉默还抑制了Akt体内激活。结论 Prx 1具有调控人肝细胞中EGFR介导信号通路的潜在作用。Prx 1能够通过EGFR-Akt信号通路介导细胞正常至异常转化。Objectives To investigate the role of Prx 1 in hepatocellular carcinoma. Methods The expression of Prx-1, pAkt and Akt in distal hepatocellular carcinoma and para- carcinoma tissue was detected by Western blot.Human liver cells were treated with 10 ng/ml EGF for 10 min after 24 hrs after serum starvation. Total proteins were extracted immediately after 10-min EGF treatment and used for Western blot. Equal amount of protein（20 μg） was separated by SDS-PAGE and probed with antibodies specific to Prx 1, pAkt, various phosphorylated forms of EGFR and total EGFR. Prx 1 protein levels in parental（untransfected）, scramble, and Prx-1shRNA transfected HepG2 cells were measured by Western blot. Reactive oxygen species（ROS） levels determined from HepG2 sublines by using probe DCFH-DA. HepG2/Prx 1Sc and HepG2/Prx-1KD cells were injected into the dorsal flank of male SCID mice.Tumor size was measured every week after injection. Protein extracted from HepG2/Prx 1Sc and HepG2/Prx 1KD tumor tissues was subjected to Western blot analysis. Equal amount of protein（20 μg） was separated by SDS-PAGE and probed with antibodies specific to Prx-1and pAkt. Results Increased expression of Prx 1 was observed in abnormal（pre-malignant） liver cells compared to their paired normal liver cells. Prx 1 enhances EGF-mediated EGFR and Akt activation. Prx-1 knock-down inhibits tumor growth of HepG2 xenografts in vivo. Prx 1 knock-down reduces liver metastasis of HepG2 xenografts in vivo. Prx-1 knock-down inhibits Akt activation in vivo. Conclusions A potential role for Prx-1 in regulation of EGFR mediated signaling pathways in human liver cells. Prx-1 in mediates normal-to-abnormal cell transition, tumorigenesis, and metastasis through EGFR-Akt signaling pathways.

关 键 词：过氧化物酶1 肝癌 移植瘤 

分 类 号：R735.7[医药卫生—肿瘤]