VEGF反义寡核苷酸对舌鳞癌影响的实验研究

目的:研究血管内皮细胞生长因子反义寡核苷酸(VEGF-ASODN)对裸鼠荷人舌癌肿瘤的生长抑制作用。方法:20只BALB/C裸鼠分为4组,每组5只,实验1～3组建立人舌癌荷瘤裸鼠移植瘤模型,并注射相应试剂;第4组为未荷瘤对照组。①VEGF-ASODN组:于瘤体及瘤周注射VEGF-ASODN66μg。②错义寡核苷酸组:注射错义寡核苷酸66μg。③生理盐水组:注射生理盐水。各组每3天注射1次,共注射7次。实验结束后2d,处死动物,取血清测VEGF蛋白,测量肿瘤大小及重量,取肿瘤组织切片用免疫组织化学方法检测VEGF、PCNA、CD34的表达,计算微血管密度(microvesseldensity,MVD)和增殖细胞核抗原标记指数(PCNALabelindex,PLI),分子原位杂交测VEGFmRNA表达。结果:VEGF-ASODN组荷瘤裸鼠血清VEGF蛋白水平,肿瘤的体积和重量,VEGFmRNA及其蛋白和PCNA、CD34的表达均显著低于对照组。结论:VEGF-ASODN通过抑制VEGF表达,从而抑制舌癌细胞的生长、增殖。

Inhibitory Effect of VEGF Antisense Phosphorothioate Oligodeoxynucleotides on the Growth of Human Tongue Cancer Xenografts in Nude Mice

Objective： To observe the inhibitory effect of VEGF （vascular endotheial growth factor） antisense phosphorothioate （VEGF-ASODN） oligodeoxynucleoiides on the proliferation of human tongue cancer xenografts. Methods： The VEGF-ASODN was artificially synthesised. After the model of human tongue cancer xenografis in 15 nude mice were established, animals were randomly divided into 3 groups： antisense group, scrambled group, saline group. 5 nude mice without tumor were used as control group. Antisense （66 μg）,scrambled sequence （66 μg） and saline were injected in three experiment groups （once even）., three days and 7 times in all）. Two days after the injection, the mice were sacrificed. Serums were used for detection of VEGF protein. All tumors were measured and weighted. The quantity of VEGFmRNA and protein, PLI and MVD were detected by hybridization in situ and immunohistochemistry. Results： In experiment group 1, serum VEGF protein level, tumor weight and volume, expressions of VEGFmRNA, PCNA,CD34 were significantly lower than that of the control group. Conclusion： By inhabiting the expression of VEGF, VEGF-ASODN can inhabit proliferation of human tongue cancer xenografis in nude mice.