HOXB2反义寡核苷酸对原代脐静脉内皮细胞生物学特性的影响

目的：探讨同源盒基因B2（HOXB2）反义硫代寡核苷酸对原代脐静脉内皮细胞生物学特性的影响。方法：采用荧光标记观察HOXB2反义寡核苷酸（Asodn）在内皮细胞中的分布;观察不同浓度HOXB2反义寡核苷酸对内皮细胞DNA合成的影响;应用流式细胞仪和RT－PCR观察其对细胞周期及HOXB2表达的影响。结果：经脂质体介导,转染15min胞核有较弱荧光染色,4－8h胞核荧光最强,16h荧光减弱、消散;HOXB2反义寡核苷酸能抑制内皮细胞DNA的合成,表现出浓度依赖效应,能延缓内皮细胞由G1期进入S期;同时HOXB2 mRNA表达水平在24－48h明显下降。结论：HOXB2在内皮细胞增殖中起重要作用,其作用机制与细胞周期有关。

Effects of HOXB2 antisense oligodeoxynucleotides on the biological properties of primary human umbilical vein endothelial cells

OBJECTIVE: To explore the effects of HOXB2 antisense oligodeoxynuc leotides (Asodn) on the biological properties of primary human umbilical vein endothelial cells (ECs). METHODS: Fluorescent labelled Asodn was transfected into the endothelial cells of human unbilical vein mediated liposome and its distribution within endothelia was observed. (3)H-TdR incorporation test was employed to determine its effects on the DNA synthesis. Flow cytometry was applied to determine the change of the cell cycle. In the same time, RT-PCR was adopted to study the influence of Asodn on the expression of target genes. RESULTS: Fifteen minutes after the transfection, weak nucleic staining was observed. The fluorescent staining was the strongest 4 approximately 8 hours after the transfection and began to weaken in 16 hours. The proportion of cells in G1/0 phase in Asodn group was 53.4 +/- 3.1, significantly higher than that in control group (35.8 +/- 7.3, P < 0.01), and the proportion of cells in S phase in Asodn group was 42.2 +/- 3.5, significantly lower than that in control group (60.8 +/- 6.2, P < 0.01). The expression of HOXB2 mRNA was remarkably decreased during 24 to 48 hours. CONCLUSION: HOXB2 Asodn exerts inhibitory effects on EC proliferation dose-dependently, delays the conversion of G1 phase to S Phase, and inhibits the expression of HOXB2 mRNA. HOXB2 gene plays an important role in proliferation of endothelial cells and the mechanism is related to cell cycle.