反义基因转染对人膀胱癌细胞血管内皮生长因子表达的影响

Effect of antisense VEGF gene transfection on the expression of VEGF in human bladder cancer cells

目的观察以腺相关病毒为载体(rAAV)的血管内皮细胞生长因子(VEGF)反义基因对人膀胱癌细胞中VEGF表达的影响。方法通过用不同量(0、20、100μl)的以腺相关病毒为基因载体的反义VEGF基因转染人膀胱癌细胞T24,采用免疫组织化学和Western blot方法,检测转染前后人膀胱癌T24细胞中VEGF的表达变化。结果免疫组织化学图像分析VEGF平均灰度值分别为364．40±30．31、460．25±36．34、494．18±38．82,Western blot蛋白条带灰度分别为 65 800±13 824、52 470±10 589、31 069±8 642。结果均表明随着转染的反义VEGF基因量的增多, T24细胞中VEGF的表达显著减少。结论反义VEGF基因的转染能显著抑制人膀胱癌T24细胞中VEGF的表达,有望成为膀胱肿瘤基因治疗的一种新方法。

Objective To study the effect of antisense VEGF gene transfection on the expression of VEGF in human bladder cancer cells with the help of adeno-associated virus-mediated vector （rAAV） and explore a new gene therapy strategy for bladder tumor. Methods Different doses （0, 20,100 μl） of antisense VEGF genes were transferred into human bladder cancer T24 cells by rAAV. Immunohistochemical and Western blot techniques were used to detect the expression of VEGF in T24 ceils. Results Immunohistochemical image analysis showed that the average gray scale values of VEGF were 364.40 ± 30.31,460.25 ± 36.34 and 94.18 ± 38.82, respectively. Western blot protein gray scale values were 65 800 ± 13 824, 52 470 ± 10 589 and 31 069 ± 8 642, respectively. The results showed that the expression of VEGF was decreased significantly with the dose increase of antisense VEGF genes transfected. Conclusion The antisense VEGF gene transfection can significantly inhibit the expression of VEGF in human bladder cancer T24 ceils and hopefully becomes an effective gene therapy approach in the treatment of bladder tumor.