TNP-470联合5-FU抑制结肠癌生长的实验研究  

Inhibitory Effects of TNP-470 in Combination with 5-Fluorouracil on Growth of Human Colon Cancer

作　　者：范应方[1] 黄宗海[1] 聂晶 

机构地区：[1]第一军医大学珠江医院普外科,广东广州510282 [2]南昌恒湖医院外科,江西南昌330123

出　　处：《癌症》2002年第12期1319-1323,共5页Chinese Journal of Cancer

基　　金：广东省自然科学基金项目(013072)

摘　　要：背景与目的:新生血管生成是肿瘤生长转移的一个重要条件,本研究通过体内外实验观察血管生成抑制剂TNP-470联合5-氟尿嘧啶(5-fluorouracil,5-FU)对结肠癌生长的影响。方法:采用四甲基偶氮唑盐(MTT)法检测TNP-470和5-FU对LOVO细胞的生长抑制效应,用Burgi法分析联合用药效果。建立结肠癌皮下移植瘤模型,观察TNP-470单独或联合5-FU对移植瘤的生长抑制作用,采用免疫组化和图像分析系统定量检测肿瘤组织中血管内皮生长因子(VEGF)的表达,抗Ⅷ因子抗体标记计数肿瘤微血管密度(MVD)。结果:TNP-470和5-FU对体外培养的LOVO细胞的生长具有抑制作用,半数抑制浓度(IC50)分别为55.8ng/ml和4.6ng/ml,Burgi法分析表明两者联合应用具有协同效应,LOVO细胞生长进一步受到抑制。体内实验表明:各治疗组肿瘤的生长明显受到抑制,而联合治疗组抗瘤作用进一步增强。联合治疗组和5-FU组肿瘤组织中VEGF的表达较对照组显著减少,联合治疗组和TNP-470组肿瘤组织中的MVD与对照组比较也显著降低。结论:TNP-470能够抑制LOVO细胞及其皮下移植瘤的生长和新生血管形成,与5-FU联合应用具有协同作用,可提高抗瘤效应。Background & Objective:Neovascularization is critical for growth of primary tumors and their distant metastasis. This study was designed to investigate the effects of angiogenesis inhibitor TNP 470 in combination with 5 fluorouracil (5 FU) on the growth of human colon cancer cell line,LOVO,in vitro and in vivo. Methods:In vitro,the growth inhibitory effect of TNP 470 or 5 FU on LOVO cells was examined by MTT assay, and synergetic effect of these two agents was evaluated by Burgi analysis. In vivo, human colon cancer xenografts were transplanted into BALB/C nude mice and treated with TNP 470 alone and combination with 5 FU. Tumor microvessel density (MVD) was measured by immunostaining with anti von Willebrand factor monoclonal antibody;the expression of vascular endothelial growth factor(VEGF) in tumor tissue was detected using an immunohistochemical method with image analysis system. Results:In vitro, TNP 470 and 5 FU inhibited the growth of LOVO cells, with the IC50 at 55.8 ng/ml and 4.6 ng/ml, respectively. Burgi analysis revealed synergism of these two agents, a remarkable growth inhibitory effect on LOVO cells was obtained. In vivo, in every treatment group,tumor growth was suppressed significantly. The combination group showed significant enhancement in anti tumor efficacy.Expression of VEGF in tumors was clearly inhibited by TNP 470 in combination with 5 FU or 5 FU alone compared to the control. Tumor MVD was lower in TNP 470 combination with 5 FU group or TNP 470 group compared to control group. Conclusion:TNP 470 suppressed the growth of LOVO cells and human colon cancer xenografts and the angiogenesis; TNP 470 in combination with 5 FU might product synergetic effect.

关 键 词：实验研究 TNP-470 5-FU LOVO细胞 结肠癌 血管生成 免疫组化 四甲基偶氮唑盐法 

分 类 号：R735.35[医药卫生—肿瘤]