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目的:探讨氨氯吡咪(Amiloride)通过抑制血管内皮细胞生长因子mRNA(VEGF mRNA)表达,观察VEGF mRNA与K562细胞凋亡是否有关。方法:将不同浓度Amiloride与K562细胞共同孵育,采用Annexin VFITC/PI双标记流式细胞术检测细胞凋亡。同时采用RT-PCR技术观察Amiloride对K562细胞VEGF mRNA表达的影响,免疫组化方法观察Amiloride对K562细胞VEGF蛋白表达的影响。应用MTT法观测Amiloride对K562细胞增殖的影响。结果:不同浓度Amiloride作用K562细胞24h后,K562细胞的增殖受到抑制,VEGF mRNA表达强度的相对系数及VEGF蛋白呈浓度依赖性下降,并且可以观察到随着VEGF mRNA表达强度的相对系数的降低,K562细胞凋亡率增加,二者之间显著负相关(P<0.05)。结论:氨氯吡咪可能通过抑制VEGF mRNA表达促进K562细胞凋亡。
AIM: To investigate whether amiloride inhibits the expression of vascular endothelial growth factor mRNA (VEGF mRNA) and whether VEGF mRNA is involved in the apoptosis of K562 cells. Methods: Different concentrations of Amiloride and K562 cells were co-incubated with Annexin VFITC / PI double-labeled flow cytometry to detect apoptosis. Meanwhile, the effect of Amiloride on VEGF mRNA expression in K562 cells was observed by RT-PCR. The effect of Amiloride on VEGF protein expression in K562 cells was observed by immunohistochemistry. The effect of Amiloride on the proliferation of K562 cells was observed by MTT assay. Results: After treated with different concentrations of Amiloride, the proliferation of K562 cells was inhibited. The relative expression of VEGF mRNA and VEGF protein decreased in a concentration-dependent manner. With the decrease of the relative expression of VEGF mRNA, K562 cells apoptosis rate increased significantly negative correlation (P <0.05). Conclusion: Aminopyram may promote the apoptosis of K562 cells by inhibiting the expression of VEGF mRNA.