摘要
目的:研究小干扰RNA(siRNA)对血管内皮细胞Akt1基因表达的沉默作用及其对肺癌A549细胞条件培养基促血管内皮细胞迁移的影响。方法:以人脐静脉内皮细胞为靶细胞,设计针对Akt1基因mRNA的siRNA(siAkt1),利用阳离子脂质体介导转染人脐静脉内皮细胞,采用半定量PCR技术(RT-PCR)检测Akt1mRNA的表达,用Western blotting技术检测Akt总蛋白表达。同时用体外"伤口愈合"实验观察细胞迁移。结果:筛选出的siAkt1能显著抑制内皮细胞Akt1mRNA(P<0.01)和Akt总蛋白的表达(P<0.05),A549细胞条件培养基显著促进人脐静脉内皮细胞迁移(P<0.05),但对转染siAkt1的人脐静脉内皮细胞迁移没有促进作用(P>0.05)。结论:特异性siRNA通过有效抑制Akt1基因的表达而抑制A549细胞条件培养基促血管内皮细胞迁移的作用,这为肺癌血管生成干预治疗提供了新的理论基础。
Objective To investigate the effect of siRNA on Aktl gene expression in endothelial cells, and its influence on the migration of endothelial cells induced with lung carcinoma A549 cells conditioned medium （CM）. Methods Using human umbilical vein endothelial cells （HUVECs） as the target cells, a siRNA sequence targeted Aktl gene（ siAktl ） was designed and transfected into HUVECs mediated with liposome. The expression of Aktl mRNA was examined with RT-PCR, and the Akt protein expression was determined with Western blot. The migration of HUVECs was obsereved with external wound healing experiment. Results HUVECs transfected with siAktl could markedly inhibited the expression of Aktl mRNA （ P 〈 0.01 ） and Akt protein （ P 〈 0. 05 ） in endothelial cells ,the CM could significantly stimulated the migration of HUVECs （P 〈 0.05）. However, CM had no significant effect on the migration of HUVECs transfected with siAktl （P 〉0.05 ）. Conclusion Specific siRNA could inhibit the migration of HUVECs induced with A549 cells CM, which may provide a novel strategy for inhibiting tumor angiogenesis.
出处
《郧阳医学院学报》
CAS
2009年第2期105-108,共4页
Journal of Yunyang Medical College
基金
湖北省教育厅重点项目(B200524009)
