大黄素抑制人肝癌HepG2细胞血管生成作用及其机制研究  

Effect of emodin on angiogenesis and its mechanism in human hepatoma cell line HepG2

作　　者：高俊霞[1] 刘爽[1] 任公平[2] 吕强 

机构地区：[1]牡丹江医学院红旗医院急诊科,黑龙江牡丹江157011 [2]牡丹江医学院红旗医院中西医结合科,黑龙江牡丹江157011 [3]穆棱市八面通林业局职工医院CT室,黑龙江穆棱157500

出　　处：《中华肿瘤防治杂志》2016年第3期164-170,共7页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的大黄素可促进人肝癌HepG2细胞的凋亡,然而其是否可抑制肝细胞癌(hepatocellular carcinoma,HCC)血管生成及其机制罕见报道。本研究探讨大黄素对人肝癌HepG2细胞血管生成以及缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的影响,以及大黄素体内抗肝癌机制。方法采用体内鸡胚绒毛尿囊膜(chick chorioallantoic membrane,CAM)实验,实验随机分为阴性对照组(生理盐水)、大黄素低剂量组(10μmol/L)、大黄素高剂量(20μmol/L)和阳性对照组(0.15mg/mL地塞米松),每组各10只,每24h每组追加同样等量药物,共72h,观察大黄素对CAM血管生成的抑制作用。体外培养HepG2细胞,以氯化钴(CoCl2)模拟化学缺氧,设立缺氧未处理组〔阴性对照组(生理盐水)〕、缺氧大黄素低剂量组(10μmol/L)、缺氧大黄素高剂量组(20μmol/L)和缺氧阳性对照组〔10μmol/L 5-氟尿嘧啶(5-fluorouracil,5-FU)〕,每组设6个复孔,处理24h。采用小管形成实验,观察大黄素对HepG2细胞相对小管数目的影响,免疫细胞化学分析检测HIF-1α和VEGF阳性细胞的表达,Real-Time PCR分析检测HIF-1αmRNA和VEGF mRNA的表达。采用Graphpad 5.0软件进行描述性统计,组间比较采用one-way-ANOWA和Bonferroni检验。结果 CAM实验显示,实验组新生血管数目明显减少,与阴性照组比较,差异有统计学意义,F=13.374,P=0.002。阴性对照组新生血管数目为(31.47±1.81)个。给药后CAM血管生成减少,大黄素低剂量组、高剂量组和地塞米松组分别为(19.48±0.66)、(10.33±1.04)和(9.89±0.57)个,与阴性对照组比较,差异有统计学意义,P值分别为0.041、0.004和0.003;大黄素低剂量组和高剂量组与地塞米松组比较,差异无统计学意义,P值分别为0.539和1.000。小管形成实验显示,实验组相对小管数目明显减少,与阴性照组比较,差异有统计学意义,F=21.529,P<0.001。阴性对照组相对小ORJECTIVE Emodin can promote the apoptosis of the human hepatoma HepG2 cells, however,whether it inhibits hepatocellular careinoma（HCC）of angiogenesis and its mechanism has not been reported. The purpose of this study is to investigate the effect of emodin on angiogenesis and hypoxia inducible factor-1α （HIF-1a） and vascular endothelial growth factor （VEGF）in human hepatoma HepG2 cells,and discuss its possible anti-cancer mechanism. METHODS The chick chorioallantoie membrane （CAM） experiment was randomly divided into the negative control group （saline）,the low dose group （10 μmol/L）, the high dose group（ 10 μmol/L） and the positive control group（0. 15 mg/mL dexamethasone）, each group had 10 CAMs, the drug was administered respectively for 24 h,a total of 72 h,the inhibition of emodin on CAM angiogenesis was observed. HepG2 cells were cultured in vitro, cobalt chloride（CoCl2 ） was added in the medium to simulate the chemical hypoxia environment, the hypoxia untreated group was established [the negative control group（saline）J, the hypoxia low dose group （10μmol/L）, the hypoxia high close group （20μmol/L） ,and the hypoxia positive control group [10 μmol/L 5-fluorouracil（5-FU）], and each group set up 6 wells,processing 24 h. The effect of emodin on the number of relatively small tube was observed in HepG2 cells by tube formation assay,the expression of HIF-1α and VEGF positive cells was detected by immunocytochehmical analysis, the expression of HIF-1α mRNA and VEGF mRNA was detected by Real-Time PCR analysis. The Graphpad 5.0 software was used in statistics, One-way-ANOWA and Borderroni test was used to compare between the groups. RESULTS The CAM experimental results showed that the number of new blood vessels in the experimental group was significantly reduced, compared with the negative control group, the difference was statistically significant （F= 13. 374, P = 0. 002）. The number of new blood vessels of the negative control group was（31.47± 1.81�

关 键 词：大黄素 肝肿瘤 血管生成 缺氧诱导因子-1Α 血管内皮生长因子 

分 类 号：R735.7[医药卫生—肿瘤]