中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (46): 8667-8670.doi: 10.3969/j.issn.2095-4344.2012.46.023

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

siRNA沉默FOXO3a对三氧化二砷抑制内皮细胞增殖的影响

李海莉，柳 红   

  1. 徐州医学院病理学教研室，江苏省徐州市 221002
  • 收稿日期:2012-01-14 修回日期:2012-03-26 出版日期:2012-11-11 发布日期:2012-11-11
  • 通讯作者: 柳红，教授，硕士生导师，徐州医学院病理学教研室，江苏省徐州市 221002
  • 作者简介:李海莉★，女，1987年生，山东省泰安市人，汉族，徐州医学院在读硕士，主要从事肿瘤病理学研究。 hailey2012@foxmail.com

Small interfering RNA mediated silencing of FOXO3a influences the arsenic trioxide-inhibited proliferation of endothelial cells

Li Hai-li, Liu Hong   

  1. Department of Pathology, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China
  • Received:2012-01-14 Revised:2012-03-26 Online:2012-11-11 Published:2012-11-11
  • Contact: Liu Hong, Professor, Master’s supervisor, Department of Pathology, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China
  • About author:Li Hai-li★, Studying for master’s degree, Department of Pathology, Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China hailey2012@foxmail.com

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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4 μmol/L As2O3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4 μmol/L As2O3的RPMI-1640培养基中培养。③FOXO3a siRNA组：转染FOXO3a siRNA后，细胞在含4 μmol/L As2O3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。

关键词: 人脐静脉内皮细胞, As2O3, FOXO3a, 血管内皮生长因子, 基因转染

Abstract:

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 4 μmol/L As2O3 after adherent. (2)In control small interfering RNA group, human umbilical vein endothelial cells were cultured in RPMI-1640 medium containing 4 μmol/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 4 μmol/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.

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