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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 observe the effect of vascular endothelial growth factor (VEGF) antisense gene with adeno-associated virus vector (rAAV) on the expression of VEGF in human bladder cancer cells. Methods Human bladder cancer cell line T24 was transfected with antisense VEGF gene with different amounts of adeno-associated virus (0, 20, and 100μl). Immunohistochemistry and Western blot were used to detect the expression of T24 Changes in the expression of VEGF in cells. Results The average gray value of VEGF in immunohistochemistry was 364.40 ± 30.31, 460.25 ± 36.34 and 494.18 ± 38.82, respectively. The gray level of Western blot was 65 800 ± 13 824,52 470 ± 10 589,31 069 ± 8 642. The results showed that with the transfection of antisense VEGF gene increased, T24 cells in the expression of VEGF was significantly reduced. Conclusion The transfection of antisense VEGF gene can significantly inhibit the expression of VEGF in human bladder cancer T24 cells and is expected to become a new method for gene therapy of bladder cancer.