摘要
背景:课题组前期已证实As2O3可以促进乳腺癌细胞中FOXO3a因子的表达,并抑制肿瘤细胞中血管内皮生长因子的表达,但As2O3对血管内皮细胞增殖的影响,以及对血管内皮细胞中FOXO3a、血管内皮生长因子的影响尚未证实。目的:观察siRNA沉默FOXO3a对As2O3抑制人脐静脉内皮细胞增殖及血管生成的影响。方法:实验分为4组:①As2O3组:待人脐静脉内皮细胞贴壁,在含4μmol/LAs2O3的RPMI-1640培养基中培养。②control siRNA组:转染无关序列control siRNA后,细胞在含4μmol/LAs2O3的RPMI-1640培养基中培养。③FOXO3a siRNA组:转染FOXO3a siRNA后,细胞在含4μmol/LAs2O3的RPMI-1640培养基中培养。④对照组:与实验药物等体积PBS作为对照,细胞在完全培养基条件下培养。应用CCK-8方法检测细胞增殖情况,采用免疫细胞化学方法观察人脐静脉内皮细胞中FOXO3a、血管内皮生长因子蛋白的表达情况。结果与结论:与对照组相比,FOXO3a siRNA明显减弱了As2O3抑制人脐静脉内皮细胞增殖的作用,As2O3促进人脐静脉内皮细胞中FOXO3a蛋白的表达并抑制血管内皮生长因子蛋白的表达,FOXO3a siRNA处理后明显抑制FOXO3a蛋白表达且增加血管内皮生长因子蛋白表达。结果提示FOXO3a可能成为As2O3抑制肿瘤细胞增殖及血管生成治疗的重要靶点。
BACKGROUND:Early studies have confirmed that As2O3 can promote FOXO3a factor expression in breast cancer cells and inhibit vascular endothelial growth factor expression in tumor cells.However,the effect of As2O3 on proliferation,FOXO3a and growth factors in vascular endothelial cells remains unclear.OBJECTIVE:To investigate the effect of silencing of FOXO3a using small RNA on inhibiting the As2O3-inhibited proliferation of human umbilical vein endothelial cells and angiogenesis.METHODS:There were four groups in this experiment:As2O3 group,control small interfering RNA group,FOXO3a small interfering RNA group and control group.(1)In As2O3 group,human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4μmol/L As2O3 after adherent.(2)In control small interfering RNA group,human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4μmol/L As2O3 after irrelevant sequence small interfering RNA was tranfected.(3)In FOXO3a small interfering RNA group,human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4μmol/L As2O3 after FOXO3a specific small interfering RNA was transfected into the cells.(4)In control group,a volume of PBS equal to the original volume of As2O3 served as a control,and human umbilical vein endothelial cells were cultured in the complete medium.After that,cell proliferation was detected by CCK-8 kit;the expressions of FOXO3a protein and vascular endothelial cell growth factor proteins in human umbilical vein endothelial cells were observed by immunocytochemical method.RESULTS AND CONCLUSION:Compared with the control group,the inhibitory effect of As2O3 on the proliferation of human umbilical vein endothelial cells was weakened by silencing of FOXO3a.Besides,As2O3 could promote FOXO3a protein expression and inhibit vascular endothelial cell expression.However,FOXO3a protein expression was inhibited obviously and protein expression of vascular endothelial growth factor was increased after FOXO3a using small interfering RNA.These results suggest that the transcription factor of FOXO3a can be an important strategy for the inhibition of tumor cell proliferation and angiogenesis.
出处
《中国组织工程研究》
CAS
CSCD
2012年第46期8667-8670,共4页
Chinese Journal of Tissue Engineering Research