EPCs在肝癌模型中的分布及其对肝癌形成的影响  

Distribution of exogenous endothelial progenitor cells in a rat model of hepatoma and their impact on liver cancer formation

作　　者：郭宁刚[1] 邵泽勇[2] 吕锋 

机构地区：[1]四川省雅安市人民医院中西医结合科,四川省雅安市625000 [2]四川省雅安市人民医院消化内科,四川省雅安市625000 [3]江苏省兴化市人民医院,江苏省兴化市225700

出　　处：《世界华人消化杂志》2011年第16期1666-1673,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30600727~~

摘　　要：目的:探讨内皮祖细胞(EPCs)在大鼠肝癌模型形成中的作用.方法:分离培养大鼠EPCs,体外用4',6-二脒基-2-苯基吲哚(DAPI)标记后,将EPCs-DAPI制成1×105/150μL,2×105/150μL,4×105/150μL的低、中、高浓度悬液,模型组和EPCs移植组的SD大鼠腹腔注射二乙基亚硝胺(DEN)制成肝癌模型,空白组注射等量的生理盐水,EPCs移植组经尾静脉注射低、中、高浓度EPCs-DAPI悬液1mL,空白组和模型组经尾静脉注射生理盐水1mL,分别于第4、6、8周末处死,留取血清和肝脏组织标本,检测EPCs标志物CD34、CD133和KDR表达,检测各组血清ALT、AST、AFP的表达,观察EPCs-DAPI表达的量及分布的变化,并用Imageproplus图像分析软件对EPCs-DAPI表达的阳性区域面积(PA)作半定量分析.结果:培养的细胞7d后检测细胞表面标志物CD34、CD133和KDR的表达均呈阳性,低浓度EPCs移植4wk组较ALT、AST、AFP模型对照4wk组显著升高(P<0.01);中浓度EPCs移植6wk组较ALT、AST、AFP模型对照6wk组显著升高(P<0.01);高浓度EPCs移植8wk组较ALT、AST、AFP模型对照8wk组显著升高(P<0.01);EPCs移植组中DAPI均呈阳性,且依注射EPCs悬液浓度高低和时间长短呈上升趋势,表达逐渐增强,在视野中密集分布,染色渐深.EPCs低、中、高浓度移植4wk组,EPCs低、中、高浓度移植6wk组和EPCs低、中、高浓度移植8wk组PA值比较,差异具有显著性(P<0.05);低浓度EPCs移植4、6、8wk组,中浓度EPCs移植4、6、8wk组和高浓度EPCs移植4、6、8wk组PA值比较,差异无显著性.结论:EPCs在肝癌形成过程中对肝功能有影响,且随着造模时间的延长而加重.EPCs在肝癌形成过程中有促进作用.EPCs体内示踪在肝组织的分布和表达水平表明其对肝癌有靶向作用.AIM： To investigate the role of endothelial progenitor cells （EPCs） in the formation of liver cancer and to explore the possibility of using EPCs as a delivery vehicle for the treatment of liver cancer. METHODS： Isolated and cultured EPCs were labeled with DAPI in vitro. Sprague-Dawley rats were divided into three groups： control group, model group, and EPC group. The model group and EPC group were intraperitoneally injected with diethylnitrosamine （DEN） to induce liver cancer, while the control group was injected with saline. Rats of the EPC group were further divid-ed into three subgroups and intravenously injected with 1 mL of low （1 × 105/150 μL）, medium （2 × 105/150 μL）, and high （4 × 105/150 μL） concentrations of EPCs-DAPI suspension, respectively, while the control group and model group were injected with equal volume of saline. At weeks 4, 6 and 8, rats were killed to take liver tissue samples to detect the expression of EPCs markers CD34, CD133, and KDR, and serum samples to measure the levels of ALT, AST, and AFP. EPCs-DAPI-positive area （PA） was quantif ied using the Imagepro plus image analysis software. RESULTS： Positive expression of cell surface markers CD34, CD133, and KDR could be detected on cultured cells after 7 days of culture. Serum ALT, AST, and AFP differed signif icantly between the low-concentration EPC group and model group at week 4, between the medium-concentration EPC group and model group at week 6, and between the high-concentration EPC group and model group at week 8 （all P 0.01）. Positive DAPI was detected in all the EPC groups, and the positive rate of DAPI was de- pendent on the concentration of injected EPCs and injection duration. PA was significantly different among rats injected with different concentrations of EPCs at all time points （all P 0.05）, but showed no significant differences among rats injected with the same concentration of EPCs at different time points （all P 0.05）. CONCLUSION： EPCs affect liver function in th

关 键 词：内皮祖细胞 肝癌 靶向治疗 载体 

分 类 号：R735.7[医药卫生—肿瘤] R-332[医药卫生—临床医学]