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目的:观察5-氮杂胞苷及转录因子Sp1(special Protein 1)抑制剂白桦脂酸(betulinic acid)对T24人膀胱癌细胞株血管内皮生长因子(vascular erndothelial growth factor,VEGF)及其异构体表达的影响,揭示VEGF基因异构体表达的变化规律。方法:分别以不同浓度的5-氮杂胞苷、白桦脂酸作用于T24细胞,RT-PCR检测细胞内总VEGF、刺激性异构体VEGF_(165)和VEGF_(121)、抑制性异构体VEGF_(165)b的mRNA表达变化;构建VEGF_(165)b表达质粒稳定转染T24细胞,RT-PCR检测外源性VEGF_(165)b对VEGF_(165)表达的影响。结果:5-氮杂胞苷作用T24细胞后,细胞总VEGF mRNA及其刺激性异构体VEGF_(165)和VEGF_(121),表达明显增加,而抑制性异构体VEGF_(165)b表达减少;以白桦脂酸作用T24细胞后,细胞内总VEGF及刺激性异构体表达减少,但抑制性异构体VEGF_(165)b表达增加;而稳定转染VEGF_(165)b表达质粒的T24细胞,VEGF_(165)表达减少。结论:人膀胱癌细胞株T24受干预后,VEGF及其异构体基因的变化有一定规律。当细胞内总VEGF表达增加时,刺激性异构体表达增加而抑制性异构体表达减少;而总VEGF表达减少时,刺激性异构体表达减少而抑制性异构体表达增加。此外,VEGF_(165)b可拮抗VEGF_(165)的表达。
AIM: To investigate the effects of 5-azacytidine and betulinic acid, a Sp1 inhibitor, on the expression of vascular endothelial growth factor (VEGF) and its isoforms in T24 human bladder cancer cell lines Body expression, revealed the variation of VEGF gene isoform expression. Methods: T24 cells were treated with different concentrations of 5-azacytidine and betulinic acid respectively. The total cellular VEGF, the stimulatory isoforms VEGF_ (165) and VEGF_ (121), inhibitory isoforms The expression of VEGF 165 B was detected by RT-PCR. The expression of VEGF 165 B was stably transfected into T24 cells, and the effect of exogenous VEGF 165 b on the expression of VEGF 165 was analyzed by RT-PCR. Results: After treated with 5-azacytidine, the expression of VEGF mRNA and its stimulatory isoforms VEGF 165 and VEGF 121 were significantly increased, while the inhibitory isoform VEGF 165b was expressed Reduced the expression of VEGF and (165) b in T24 cells, while the expression of VEGF165 (b) increased in the cells. However, the expression of VEGF165 T24 cells, VEGF_ (165) expression decreased. Conclusion: The changes of VEGF and its isoforms in human bladder cancer cell line T24 are regulated by some rules. When total intracellular VEGF expression increased, the expression of stimulatory isoform increased and the expression of inhibitory isoform decreased. However, when the total VEGF expression decreased, the expression of stimulating isoform decreased and the expression of inhibitory isoform increased. In addition, VEGF_ (165) b can antagonize the expression of VEGF_ (165).