血管内皮细胞生长因子反义寡核苷酸联合低分子肝素治疗小鼠Lewis肺癌的研究

目的探讨血管内皮细胞生长因子（VEGF）反义寡核苷酸和低分子肝素法安明单用或联用对小鼠Lewis肺癌生长、转移的抑制作用。方法复制小鼠Lewis肺癌模型40只,随机分为对照组、VEGF反义寡核苷酸（ASODN）组、VEGF错义寡核苷酸（MSODN）组、低分子肝素（LMWH）组及联合治疗组,每组8只,分别给予生理盐水、VEGF-ASODN、VEGF-MSODN、法安明及VEGF-ASODN＋法安明皮下注射,隔日1次,共15次。检测皮下移植瘤变化和肺转移率;应用免疫组织化学的方法检测肿瘤组织微血管密度（MVD）;应用Western印迹的方法检查肿瘤组织VEGF蛋白表达。结果ASODN、LMWH和联合治疗组的抑瘤率分别为47．34％、27．31％和59．03％;肺转移率分别为37．5％、37．5％和25％;MVD和VEGF蛋白表达降低;与对照组和MSODN组相比,抑瘤率和MVD差异均有统计学意义（P〈0．05）。结论VEGF-ASODN和法安明具有下调VEGF表达,抗血管生成作用;二者联合有协同抗肿瘤作用。

The effect of VEGF antisense oligonucleotides combined with low molecular weight heparin on the growth and metastasis of mice Lewis lung cancer

Objective To investigate the inhibitory effect of vascular endothelial growth factor (VEGF) antisense oligonucleotides or/and dalteparin sodium( fragmin) on tumor growth and metastasis of mice Lewis lung cancer. Methods 40 mice with Lewis lung cancer were randomizedly divided into five groups: control group,VEGF antisense oligonucleotides (ASODN)group, VEGF mismatch sense oligonucleotides(MSODN)group, low molecular weight heparin (LMWH )group, and combined group. Sodium chloride, VEGF-ASODN, VEGF -MSODN, fragmin, and VEGF-ASODN plus fragmin were given respectively(once every two days,15 times altogether). The volume and weight of subcutaneous tumors were measured, and the rates of lung metastasis were detected by HE staining . The microvessel density (MVD) in tumor mass were measured by imuunohistochemistry staining. VEGF protein level in tumor tissue were detected by Western blot assay . Results After treatment , the tumor growth inhibitory rates were 47.34%,27.31% and 59.03%, and the rates of lung metastasis were 37.5%, 37.5% and 25% in ASODN , LMWH , and the combined group , respectively.Tumor MVD and VEGF protein expression of the above three groups were lower than those of the control group. There was a significant difference in regard to the tumor growth inhibitory rates and MVD between the above three treated groups and the control group as well as the MSODN group (P<0.05). Conclusions VEGF-ASODN and fragmin may down-regulate VEGF gene expression and inhibit angiogenesis, Combined use of fragmin can enhance anti- tumor effect of VEGF-ASODN.