摘要
目的 研究人肝癌细胞系 SMMC772 1细胞对血管内皮生长因子 (VEGF)反义寡核苷酸 (ASODN)的摄取及其分布、作用特点 ,为 ASODN的进一步应用提供相关的理论依据 .方法 采用人工合成荧光标记的 VEGF ASODN作用于SMMC772 1细胞 ,通过激光共聚焦显微镜观察 ASODN在肝癌细胞中的分布情况 ,定量分析 ASODN作用的浓度及时间特点 ,确定 ASODN作用的最佳浓度及时间 .结果 ASODN在肝癌细胞中的浓度随作用时间的延长而逐渐升高 ,12 h内即可达到作用浓度 ,4 8h达到最高峰 ,以后则逐渐降低 ,可持续 72 h以上 . ASODN在细胞内的分布密度随作用浓度的升高而升高 ,但当达到一定的作用浓度后 ,细胞内 ASODN的含量随作用浓度增加升高的并不明显 ,其中 5 μmol· L- 1是ASODN较为理想的作用浓度 .在非分裂期肝癌细胞内 ,ASODN的分布部位主要位于胞质 ,胞核也有少量分布 ,而在分裂期肝癌细胞则主要分布在胞核 ,且其对 ASODN的摄入能力远大于非分裂期细胞 .结论 对 VEGF ASODN的深入研究可使其有望成为抗肝癌的新一代基因治疗药物 .
AIM To acquire academic basis for clinical appli cation of antisense oligodeoxynucleotide (ASODN) according to investigation uptaken, distribution and effect of ASODN of vascular endothelial growth factor (VEGF) in SMMC7721 (human hepatocellular carcinoma cell line). METHODS Synthetic fluorescein labeled ASODN was added into the medium of SMMC7721 cell. Distribution of ASODN in SMMC7721 cell was observed by laser scanning confocal microscopy. In order to confirm the optimal effective concentration and process time of ASODN, the effect rule of ASODN following its concentration and process time was analyzed quantitatively. RESULTS Density of ASODN in SMMC7721 cell was correlated with process time. The effective density was achieved in 12 h, the maximal density in 48 h, and then declined gradually. The effect of ASODN was able to last for more than 72 h. The higher of the concentration of ASODN was, the higher of the density of ASODN in SMMC7721 cell was. But this was not absolute. When the concentration was 5 μmol·L -1 , the effect of ASODN was perfect. ASODN was predominantly localized to cytoplasm in SMMC7721 cells which were in interkinesis while nucleolus in fission SMMC7721 cells. The englobing capacity of SMMC7721 to ASODN was stronger in fission phage than in interkinesis. CONCLUSION It is promising that ASODN of VEGF could become a new gene of therapeutic drug for human hepatocellular carcinoma with research in depth.
出处
《第四军医大学学报》
北大核心
2002年第21期1944-1947,共4页
Journal of the Fourth Military Medical University
