血管内皮生长因子反义寡核苷酸对裸鼠淋巴瘤血管生成的影响  

Effect of Vascular Endothelial Growth Factor Antisense Oligodeoxynucleotides on Angiogenesis in Lymphoma of Nude Mice

作　　者：童允洁[1] 张敏[1] 邹萍[1] 郭荣[1] 袁永辉[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液科,湖北武汉430022

出　　处：《癌症》2005年第1期62-66,共5页Chinese Journal of Cancer

摘　　要：背景与目的:研究表明,血管内皮生长因子(vascular endothelial growth factor,VEGF)与淋巴瘤从低度向高度恶性的进展有关,且VEGF表达高的淋巴瘤易发生早期远处转移。目前,VEGF反义脱氧寡核苷酸(antisense oligodeoxynucleotides,ASODN)对淋巴瘤血管生成影响的研究尚少。本课题拟通过体内、体外实验探讨硫代磷酸化修饰的VEGFASODN对淋巴瘤血管生成的影响。方法:将终浓度分别为10、20、30μmol/L的VEGFASODN和错义序列与人淋巴瘤细胞系Namalwa细胞分别孵育24h、48h,采用RT-PCR检测VEGFmRNA的表达,采用链酶菌抗生素蛋白鄄过氧化酶免疫组化法(streptavidin/peroxidase,SP法)检测VEGF蛋白的表达。用VEGFASODN处理的Namalwa细胞注射入裸鼠,监测裸鼠肿瘤的大小,并采用SP法分析淋巴瘤中微血管密度(microvesseldensity,MVD)。结果:凝胶图像分析表明VEGFASODN3个浓度组(10、20、30μmol/L)处理的Namalwa细胞VEGFmRNA的表达分别为1.28、0.86和0.47,错义序列组和对照组分别为1.79和1.84。VEGFASODN组、错义序列组和对照组VEGF蛋白的表达分别为23.3%、46.9%和47.8%。VEGFASODN组、错义组、PBS组的细胞接种裸鼠,其新生血管的数目分别为12.26±0.78、23.92±1.14和24.13±1.21。BACKGROUND & OBJECTIVE: It has been reported that vascular endothelial growth factor (VEGF) plays an important role in progression of lymphoma, and lymphoma with high level of VEGF is likely to metastasise at early stage. This study was to explore effects of antisense oligodeoxynucleotides (ASODN) targeting VEGF on angiogenesis of lymphoma through in vitro, and in vivo experiments. METHODS: Human lymphoma cell line Namalwa cells were incubated with VEGF ASODN (final concentrations were 10, 20, and 30 μmol/L, respectively), or scrambled sequence for 24 h or 48 h. VEGF mRNA expression in treated cells was detected by reverse transcriptase- polymerase chain reaction (RT-PCR), and its protein expression was detected by SP immunohistochemistry. After incubation of 30 μmol/L of VEGF ASODN, 30 μmol/L of scrambled sequence, and PBS as negative control, Namalwa cells were injected into nude mice. Diameters of tumors in ASODN group (6 mice), scrambled sequence group (6 mice), and PBS group (4 mice) were measured, microvessel density (MVD) was detected via immunohistochemistry. RESULTS: Expression of VEGF mRNA in cells treated with 10, 20, and 30 μmol/L of ASODN were 1.28, 0.86, and 0.47, respectively; those of PBS-treated cells, and scrambled sequence-treated cells were 1.79, and 1.84. Positive rate of VEGF protein in ASODN-treated cells was significantly lower than those in scrambled sequence-treated cells, and PBS-treated cells (23.3% vs. 46.9%, and 47.8%, P<0.05). MVDs of ASODN group, scrambled sequence group, and PBS group were 12.26±0.78, 23.92±1.14, and 24.13±1.21, respectively. CONCLUSION: VEGF ASODN can down-regulate expression of VEGF, and suppress agiogenesis in lymphoma.

关 键 词：VEGF 反义脱氧寡核苷酸 Namalwa细胞 淋巴瘤 

分 类 号：R733.1[医药卫生—肿瘤]